Mixture for Transdermal Delivery of Low and High Molecular Weight Compounds

ABSTRACT

Aspects of the present invention concern the discovery of a transdermal delivery composition that can deliver low, medium and high molecular weight pharmaceuticals and cosmetic agents. Embodiments include transdermal delivery compositions with therapeutic and cosmetic application, transdermal delivery devices for providing said transdermal delivery compositions to subjects in need thereof, and methods of making and using of the foregoing.

FIELD OF THE INVENTION

Embodiments of the present invention relate to the discovery of severalformulations of a transdermal delivery composition that delivers low andhigh molecular weight compounds, particularly drugs and cosmetic agentsto a subject. Aspects of the invention include said transdermal deliverycompositions, transdermal delivery devices for providing saidcompositions to subjects in need thereof and methods of making and usingthe foregoing.

BACKGROUND OF THE INVENTION

The skin provides a protective barrier against foreign materials andinfection. In mammals this is accomplished by forming a highly insolubleprotein and lipid structure on the surface of the comeocytes termed thecornified envelope (CE). (Downing et al., Dermatology in GeneralMedicine, Fitzpatrick, et al., eds., pp. 210-221 (1993), Ponec, M., TheKeratinocyte Handbook, Leigh, et al., eds., pp. 351-363 (1994)). The CEis composed of polar lipids, such as ceramides, sterols, and fattyacids, and a complicated network of cross-linked proteins; however, thecytoplasm of stratum corneum cells remains polar and aqueous. The CE isextremely thin (10 microns) but provides a substantial barrier. Becauseof the accessibility and large area of the skin, it has long beenconsidered a promising route for the administration of drugs, whetherdermal, regional, or systemic effects are desired.

A topical route of drug administration is sometimes desirable becausethe risks and inconvenience of parenteral treatment can be avoided; thevariable absorption and metabolism associated with oral treatment can becircumvented; drug administration can be continuous, thereby permittingthe use of pharmacologically active agents with short biologicalhalf-lives; the gastrointestinal irritation associated with manycompounds can be avoided; and cutaneous manifestations of diseases canbe treated more effectively than by systemic approaches.

Most transdermal delivery compositions achieve epidermal penetration byusing a skin penetration enhancing vehicle. Such compounds or mixturesof compounds are known in the art as “penetration enhancers” or “skinenhancers”. While many of the skin enhancers in the literature enhancetransdermal absorption, several possess certain drawbacks in that (i)some are regarded as toxic; (ii) some irritate the skin; (iii) some havea thinning effect on the skin after prolonged use; (iv) some change theintactness of the skin structure resulting in a change in thediffusability of the drug; and (v) all are incapable of delivering highmolecular weight pharmaceuticals and cosmetic agents. Despite theseefforts, there remains a need for transdermal delivery compositions thatdeliver a wide-range of pharmaceuticals and cosmetic agents.

BRIEF SUMMARY OF THE INVENTION

Disclosed herein are formulations of transdermal delivery compositionsused to deliver pharmaceuticals, therapeutic compounds, diagnostics, andcosmetic agents of various molecular weights. In several embodiments,the transdermal delivery composition comprises a unique formulation ofpenetration enhancer (an ethoxylated lipid, modified lipid, fatty acid,fatty alcohol, or fatty amine therein having 10-19 ethoxylations permolecule) or transdermal delivery enhancer (an ethoxylated compound witha multi-functional backbone) that delivers a wide range ofpharmaceuticals and cosmetic agents having molecular weights of lessthan 100 daltons to greater than 500,000 daltons. For example,embodiments of the transdermal delivery composition include formulationsthat deliver a therapeutically effective amount of a pharmaceutical,including NSAIDs, capsaicin or Boswellin-containing pain-reliefsolutions, other drugs or chemicals, dyes, low and high molecular weightpeptides (e.g., collagens or fragments thereof), hormones, nucleicacids, antibiotics, vaccine preparations, and immunogenic preparations.Methods of making the transdermal delivery compositions described hereinand systems for their delivery are embodiments. Further embodimentsinclude methods of using said compositions (e.g., the treatment andprevention of undesired human conditions or diseases or cosmeticapplications).

Aspects of the invention concern transdermal delivery compositions thatcomprise lipospheres. In some embodiments, the liposphere comprises anethoxylated composition having a carbon chain length of at least 10,wherein the ethoxylated composition (e.g., a fatty acid, fatty alcohol,or fatty amine), comprises, consists of, or consists essentially of atleast 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 ethoxylations permolecule. Aspects of the invention also include propoxylatedcompositions or compositions that comprise a combination of propoxylatedand ethoxylated compositions. In some formulations, the ethoxylated orpropoxylated composition is a fatty moiety, such as a fatty acid (e.g.,an unsaturated fatty acid or a polyunsaturated fatty acid). In otherformulations, the fatty moiety is a fatty alcohol. In other embodiments,the liposphere comprises an ethoxylated or propoxylated oil or lipidhaving carbon chain lengths of at least 10, wherein the ethoxylated orpropoxylated oil or lipid (e.g., a nut oil, a tri-alcohol, a tri-fattyamine, a glycolipid, a sphingolipid, a glycosphingolipid, or any othermodified lipid moiety), comprises, consists of, or consists essentiallyof at least 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 ethoxylations permolecule.

In preferred embodiments, the number of ethoxylations or propoxylationsper molecule is the same as the number of carbons in the fatty moiety orlipid moiety. Desirably, the fatty moiety has a carbon chain length ofat least 10, 12, 14, 16, 18, 20, 22, or 24. The liposphere comprises ahomogeneous mixture of an ethoxylated or propoxylated fatty moiety insome embodiments, while in other embodiments, the liposphere comprises aheterogeneous mixture of an ethoxylated or propoxylated fatty moiety.

Other aspects of the invention concern transdermal delivery compositionscomprised of an ethoxylated lipid moiety, such as an oil, glycolipid,sphingolipid, or glycosphingolipid. The ethoxylated oil that can be usedin the formulations described herein can be a vegetable, nut, animal, orsynthetic oil having at least 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, ormore ethoxylations per molecule. Preferred oils include macadamia nutoil or meadowfoam (limnanthes alba). It should be understood that whenan oil is ethoxylated, one or more of the components of the oil areethoxylated (e.g., fatty acids, fatty alcohols, and/or fatty amines) andit is generally recognized in the field that an average number ofethoxylations for the oil and components is obtained and thereforeprovided. That is, the measured composition is the algebraic sum of thecompositions of the species in the mix.

Still other aspects of the invention relate to transdermal deliverycompositions comprising a delivery enhancer. As used herein, the term“transdermal delivery enhancer” refers to a molecule that comprises amulti-functional backbone having at least two reactive (R) groups. The Rgroups on the multifunctional backbone comprise a reactive hydrogen,such as —OH, COOH, amines, sulfydryl groups, and aldehydes. Thus,multifunctional backbones include trialcholols, triacids, amino acids,dipeptides, tripeptides, sugars, and other compounds such asglucosamine. At least one R group is substituted with a fatty moiety,and least one reactive group is substituted with a polyethoxy orpolyethoxy/polypropoxy group, wherein the polyethoxy or thepolyethoxy/polypropoxy group comprises between 10 and 19 ethoxy orpropoxy/ethoxy substituents, respectively.

In several formulations, the ethoxylated fatty moiety is about 0.1% togreater than 99% by weight of the transdermal delivery compositiondescribed herein.

In some embodiments of the invention, the transdermal deliverycomposition further comprises an alcohol and/or water and/or an aqueousadjuvant. In some embodiments, the aqueous adjuvant is a plant extractfrom the family of Liliaceae, such as Aloe Vera. Other embodiments ofthe invention include the transdermal delivery composition describedabove, wherein about 0/1% to 15% by weight or volume is alcohol or 0.1%to 15% is water or both, or wherein about 0.1% to 85% by weight orvolume is water or Aloe Vera or another aqueous adjuvant.

Alcohol, water, and other aqueous adjuvants are not present in someformulations of the transdermal delivery composition described herein.It has been discovered that some delivered agents (e.g., steroids) aresoluble and stable in ethoxylated oil in the absence of alcohol or waterand some delivered agents are soluble and stable in ethoxylatedoil/alcohol emulsions, ethoxylated oil/water emulsions, ethoxylatedoil/alcohol/water emulsions, and ethoxylated oil/alcohol/water/Aloe Veraemulsions. In particular, it was found that a particular Aloe Vera,alcohol, or water mixture was not essential to obtain a transdermaldelivery composition provided that an appropriately ethoxylated oil wasmixed with the delivered agent. That is, the alcohol, water, and AloeVera can be removed from the formulation by using a light oil (e.g.,macadamia nut oil) that has been ethoxylated to approximately 10-19ethoxylations/molecule, desirably 11-19 ethoxylations/molecule, moredesirably 12-18 ethoxylations/molecule, still more desirably 13-17ethoxylations/molecule, preferably 14-16 ethoxylations/molecule and mostpreferably 15 or 16 ethoxylations/molecule. For example, someethoxylated oils (e.g., macadamia nut oil comprising, consisting of orconsisting essentially of 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19ethoxylations/molecule) can deliver low and high molecular weightpeptides (e.g., collagen and fragments of collagen) or amino acids inthe absence of alcohol and Aloe Vera. Some embodiments, however, have aratio of ethoxylated lipid:alcohol:aqueous adjuvant selected from thegroup consisting of 1:1:4, 1:1:14, 3:4:3, and 1:10:25.

In still other embodiments, the transdermal delivery compositionsdescribed herein can also include fragrances, creams, bases and otheringredients that stabilize the formulation, facilitate delivery, orprotect the delivered agent from degradation (e.g., agents that inhibitDNAse, RNAse, or proteases).

The transdermal delivery compositions described herein are useful forthe delivery of a wide variety of delivered agents. In certainembodiments, the transdermal delivery composition comprises deliveredagents that are hormones. In some embodiments, the delivered agent is apeptide hormone. Non-limiting examples of peptide hormones includeoxytocin, vasopressin, melanocyte-stimulating hormone, corticortropin,lipotropin, thyrotropin, growth hormone, prolactin, luteinizing hormone,human chorionic gonadotropin, follicle stimulating hormone,corticotropin-releasing factor, gonadotropin-releasing factor,prolactin-releasing factor, prolactin-inhibiting factor, growth-hormonereleasing factor, somatostatin, thyrotropin-releasing factor, calcitoningene-related peptide, parathyroid hormone, glucagon-like peptide 1,glucose-dependent insulinotropic polypeptide, gastrin, secretin,cholecystokinin, motilin, vasoactive intestinal peptide, substance P,pancreatic polypeptide, peptide tyrosine tyrosine, neuropeptidetyrosine, amphiregulin, insulin, glucagon, placental lactogen, relaxin,angiotensin II, calctriol, atrial natriuretic peptide, melatonin, andinsulin.

In other embodiments, the delivered agent is a non-peptide hormone.Non-limiting examples of hormones that are not peptide hormones usefulin embodiments include thyroxine, triiodothyronine, calcitonin,estradiol, estrone, progesterone, testosterone, cortisol,corticosterone, aldosterone, epinephrine, norepinepherine, androstiene,or calcitriol.

Other peptides such as collagen, or fragments thereof, are deliveredagents in certain embodiments.

In additional embodiments, the delivered agent is a pharmacologicallyactive small compound. For example, in certain embodiments, thedelivered agent is an anesthetic such as articaine, procaine,tetracaine, chloroprocaine and benzocaine, novocain, mepivicaine,bupivicaine, benzocaine, or lidocaine. Analgesics are delivered agentsin other embodiments. Thus, in certain embodiments the delivered agentis tramadol hydrochloride, fentanyl, metamizole, morphine sulphate,ketorolac tromethamine, hydrocodone, oxycodone, morporine, loxoprofen,Capsaicin, or Boswellin.

Other pharmacologically active compounds that are suitable deliveredagents include non-steroidal anti-inflammatory drugs (“NSAIDs”). Thus,in embodiments of the invention the delivered agent is ibuprofen(2-(isobutylphenyl)-propionic acid); methotrexate (N-[4-(2,4 diamino6-pteridinyl-methyl]methylamino]benzoyl)-L-glutamic acid); aspirin(acetylsalicylic acid); salicylic acid; diphenhydramine(2-(diphenylmethoxy)-N,N-dimethylethylamine hydrochloride); naproxen(2-naphthaleneacetic acid, 6-methoxy-9-methyl-, sodium salt, (−));phenylbutazone (4-butyl-1,2-diphenyl-3,5-pyrazolidinedione);sulindac-(2)-5-fluoro-2-methyl-1-[[p-(methylsulfinyl)phenyl]methylene-]-1H-indene-3-aceticacid; diflunisal (2′,4′,-difluoro-4-hydroxy-3-biphenylcarboxylic acid;piroxicam (4-hydroxy-2-methyl-N-2-pyridinyl-2H-1,2-benzothiazine-2-carboxamide 1,1-dioxide, an oxicam; indomethacin(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-H-indole-3-acetic acid);meclofenamate sodium (N-(2,6-dichloro-m-tolyl) anthranilic acid, sodiumsalt, monohydrate); ketoprofen (2-(3-benzoylphenyl)-propionic acid;tolmetin sodium (sodium 1-methyl-5-(4-methylbenzoyl-1H-pyrrole-2-acetatedihydrate); diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneaticacid, monosodium salt); hydroxychloroquine sulphate(2-{[4-[(7-chloro-4-quinolyl)amino]pentyl]ethylamino}ethanol sulfate(1:1); penicillamine (3-mercapto-D-valine); flurbiprofen([1,1-b]phenyl]-4-acetic acid, 2-fluoro-alphamethyl-, (+−.)); cetodolac(1-8-diethyl-13,4,9, tetra hydropyrano-[3-4-13]indole-1-acetic acid;mefenamic acid (N-(2,3-xylyl)anthranilic acid; and diphenhydraminehydrochloride (2-diphenyl methoxy-N, N-di-methylethamine hydrochloride).

In other embodiments, the delivered agent is a steroidalanti-inflammatory compound, such as hydrocortisone, prednisolone,triamcinolone, or piroxicam.

In yet other embodiments, the delivered agent is an anti-infectiveagent. By way of example, in some embodiments, the delivered agent is anantimicrobial or antifungal agent such as amoxicillin, clavulanatepotassium, itraconazole, flucanazole, erythromycin ehtysuccinate, acetylsulfisoxazole, penicillin V, erythromycin, azithromycin, tetracycline,ciproflaxin, gentamycin sulfathiazole. In still other embodiments, thedelivered agent is an anti-viral compound, such as for exampleacyclovir, lamivudine, indinavir sulfate, stavudine, saquinavir,ritonavir or hepsysls.

In still other embodiments, the delivered agent is a nucleic acid. Insome embodiments, the nucleic acid is an oligonulcoeitde consisting ofcysteine and guanidine, e.g., a CpG molecule. In further embodiments,the nucleic acid is a polynucleotide. In some embodiments, thepolynucleotide comprises a nucleic acid sequence that is capable ofeliciting an immune response from an animal. For example, in someembodiments, the nucleic acid comprises nucleic acid sequences from HIV,influenza A virus, hepatitis C virus, hepatitis A virus, hepatitis Bvirus, hantavirus, SARS, or sequences encoding members of the Inhibitorof Apoptosis family of proteins.

Embodiments of the transdermal delivery compositions disclosed hereinalso include transdermal delivery systems that comprise adjuvants andimmunogenic compositions. Thus, some formulations of transdermaldelivery compositions comprise an immunogenic peptide or nucleic acidencoding said peptide, a vaccine, such as a DNA vaccine, polypeptidevaccine, or other vaccine, and an adjuvant, such as aluminium hydroxide,calcium phosphate, cytokines (such as, e.g., interleukin-12 (IL-12)),co-stimulatory molecules (such as, e.g., B7-1 (CD80) or B7-2 (CD86)),and haptens, such as dinitrophenyl (DNP), and the like.

In yet other embodiments, the delivered agent is an immune responsemodifier. For example, in some embodiments, the delivered immuneresponse modifier is an imidazoquinoline amine including, but notlimited to, substituted imidazoquinoline amines. For example in someembodiments the delivered agent is an amide substituted imidazoquinolineamine, a sulfonamide substituted imidazoquinoline amine, a ureasubstituted imidazoquinoline amine, an aryl ether substitutedimidazoquinoline amine, a heterocyclic ether substitutedimidazoquinoline amine, an amido ether substituted imidazoquinolineamine, a sulfonamido ether substituted imidazoquinoline amine, a ureasubstituted imidazoquinoline ether, a thioether substitutedimidazoquinoline amine, or a 6-, 7-, 8-, or 9-aryl or heteroarylsubstituted imidazoquinoline amine. In other embodiments, the deliveredagent is a tetrahydroimidazoquinoline amine such as an amide substitutedtetrahydroimidazoquinoline amine, a sulfonamide substitutedtetrahydroimidazoquinoline amine, a urea substitutedtetrahydroimidazoquinoline amine, an aryl ether substitutedtetrahydroimidazoquinoline amine, a heterocyclic ether substitutedtetrahydroimidazoquinoline amine, an amido ether substitutedtetrahydroimidazoquinoline amine, a sulfonamido ether substitutedtetrahydroimidazoquinoline amine, a urea substitutedtetrahydroimidazoquinoline ether, or a thioether substitutedtetrahydroimidazoquinoline amine. In other embodiments, the deliveredagent is an imidazopyridine amine such as an amide substitutedimidazopyridine amine, a sulfonamide substituted imidazopyridine amine,a urea substituted imidazopyridine amine, an aryl ether substitutedimidazopyridine amine, a heterocyclic ether substituted imidazopyridineamine, an amido ether substituted imidazopyridine amine, a sulfonamidoether substituted imidazopyridine amine, a urea substitutedimidazopyridine ether, or a thioether substituted imidazopyridineamines. In yet other embodiments, the delivered agent is a 1,2-bridgedimidazoquinoline amine; 6,7-fused cycloalkylimidazopyridine amine, aidazonaphthyridine amine, a tetrahydroimidazonaphthyridine amines, anoxazoloquinoline amine, a thiazoloquinoline amine; an oxazolopyridineamine, a thiazolopyridine amine, a oxazolonaphthyridine amine, athiazolonaphthyridine amine, a 1H-imidazo dimers fused to a pyridineamine, a quinoline amine, a tetrahydroquinoline amine, a naphthyridineamine, or a tetrahydronaphthyridine amine.

In further embodiments, the immune response modifier is a purinederivative, an imidazoquinoline amide derivative, a 1H-imidazopyridinederivative, a benzimidazole derivatives, a derivative of a4-aminopyrimidine fused to a five membered nitrogen containingheterocyclic ring (including adenine derivatives), a3-.beta.-D-ribofuranosylthiazolo[4,5-d]pyri-midine derivative, or a1H-imidazopyridine derivatives.

Examples of particular immune response modifier compounds useful asdelivered agents include 2-propyl[1,3]thiazolo[4,5-c]quinolin-4-amine,4-amino-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol,and4-amino-2-(ethoxymethyl)-.alpha.,.alpha.-dimethyl-6,7-,8,9-tetrahydro-1H-imidazo[4,5-c]quinoline-1-ethanol.Other examples of Immune response modifier compounds includeN-[4-(4-amino-2-butyl-1H-imidazo[4,5-c][1,5]naphthyridin-1-yl)butyl]-N′-c-yclohexylurea,2-methyl-1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyri-din-4-amine,1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyridin-4-amine-,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimet-hylethyl}methanesulfonamide,N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]quinol-in-1-yl)butyl]methanesulfonamide,2-methyl-1-[5-(methylsulfonyl)pentyl]-1H--imidazo[4,5-c]quinolin-4-amine,N-[4-(4-amino-2-propyl-1H-imidazo[4,5-c]_(q)-uinolin-1-yl)butyl]methanesulfonamide,2-butyl-1-[3-(methylsulfonyl)propyl-]-1H-imidazo[4,5-c]quinoline-4-amine,2-butyl-1-{2-[(1-methylethyl)sulfony-l]ethyl}-1H-imidazo[4,5-c]quinolin-4-amine,N-{2-[4-amino-2-(ethoxymethyl)--1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimethylethyl}-N′-cyclohexylurea,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimeth-ylethyl}cyclohexanecarboxamide,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[-4,5-c]quinolin-1-yl]ethyl}-N′-isopropylurea.Resiquimod, and4-amino-2-ethoxymethyl-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinolin-e-1-ethanol.

In certain embodiments, the delivered agent is an analgesic.Non-limiting examples of analgesiscs include tramadol hydrochloride,fentanyl, metamizole, morphine sulphate, ketorolac tromethamine,morphine, and loxoprofen sodium. In other embodiments, the deliveredagent is a migraine therapeutic, such as ergotamine, melatonin,sumatriptan, zolmitriptan, or rizatriptan.

In yet other embodiments, the delivered agent is an imaging component,such as iohexyl, technetium, TC99M, sestamibi, iomeprol, gadodiamide,oiversol, and iopromide. Diagnostic contrast components such asalsactide, americium, betazole, histamine, mannitol, metyraphone,petagastrin, phentolamine, radioactive B12, gadodiamide, gadopenteticacid, gadoteridol, perflubron are delivered agents in certainembodiments.

Another aspect of the invention concerns methods of making lipopsheresuseful for transdermal delivery of a delivered agent. In one embodiment,a liposphere is made by identifying a delivered agent for incorporationinto a liposphere and mixing said delivered agent with an ethoxylated orpropoxylated fatty moiety, ethoxylated or propoxylated lipid moiety, orethoxylated or propoxylated multifunctional backbone, wherein saidethoxylated fatty moiety, lipid moiety, or multifunctional backbone hasbetween 10 and 19 ethoxylations per molecule. In preferred embodiments,the fatty moiety, or at least one of the fatty components of the lipidmoiety or multifunctional backbone has a carbon chain length of betweenabout 10 and 24 carbon residues.

The formulations described herein are placed into a vessel that isjoined to an applicator such that the active ingredients can be easilyprovided to a subject. Applicators include, but are not limited to,roll-ons, bottles, jars, tubes, sprayer, atomizers, brushes, swabs, geldispensing devices, and other dispensing devices.

Aspects of the present invention also concern compositions comprising atransdermal delivery and a transdermal delivery device, which provides ameasured amount of said transdermal delivery system. Accordingly,desired dosages of delivered agents can be delivered to a subject inneed. An exemplary transdermal delivery device is depicted in FIGS.1A-4B.

Yet other aspects of the present invention relate to methods ofdelivering an amount of a transdermal delivery composition comprisingproviding a transdermal delivery composition within a transdermaldelivery device, wherein the device is designed to administer a measuredamount of the transdermal delivery composition and providing atransdermal delivery composition to be administered to a subject.

Several methods of using the transdermal delivery compositions are alsoembodiments. For example, one approach involves a method of reducingpain or inflammation by using a transdermal delivery composition thatcomprises an anti-inflammatory molecule (e.g., an NSAID or MSM) on asubject in need of a reduction of pain or inflammation. Monitoring thereduction in inflammation may also be desired as part of arehabilitation program.

NSAIDs and other chemotherapeutic agents have also been shown to improvethe health, welfare, or survival of subjects that have cancer orAlzheimer's disease. The tendency of these compounds to cause adverseside effects such as gastrointestinal irritation liver and kidneyproblems renders them particularly desirable transdermal deliveryagents. Accordingly, some embodiments concern methods of usingtransdermal delivery compositions that comprise delivered agents (e.g.,any one or combination of the NSAIDs disclosed above or otherchemotherapeutic agents such as fluorouracil) to treat or prevent canceror hyperproliferative cell disorders (e.g., basal cell carcinoma oractinic keratosis.) For example, a method to improve the health,welfare, or survival of a subject that has cancer or Alzheimer's diseaseor a method of treating or preventing cancer or Alzheimer's disease insaid subject can be conducted by using a transdermal deliverycomposition that comprises a COX enzyme inhibitor and providing saidtransdermal delivery composition to said subject.

Some formulations of transdermal delivery compositions can be used toreduce oxidative stress to cells, tissues and the body of a subject. Forexample, a method to improve the health, welfare, or survival of asubject that is in need of a reduction in oxidative stress to a cell,tissue, or the body as a whole involves providing to said subject atransdermal delivery composition that comprises an antioxidant such asascorbic acid, tocopherol or tocotrienol or an anti-stress compound suchas Bacocalmine (Bacopa Monniera Extract obtained from SedermaLaboratories). Methods of treating or preventing diseases or conditionsassociated with oxidative stress or vitamin deficiency and methods ofreducing an oxidative stress or a vitamin deficiency in a subject inneed thereof are also embodiments.

Other formulations of transdermal delivery composition can be used toreduce psoriasis or eczema or a related condition or can be used topromote wound healing in a subject in need thereof. By one approach, atransdermal delivery composition that comprises peptides that promotewound healing (e.g., peptides comprising the sequence LKEKK (SEQ. ID.NO:1), are provided to a subject in need of a treatment or reduction inpsoriasis or eczema or a condition associated with psoriasis or eczema(e.g., allergies) or treatment of a wound.

An exemplary formulation for the treatment of Psoriasis, or Eczema is asfollows:

Hepsyl 2 Grams Distilled Water with Sodium Bi Carbonate 18 mls (to alterph to 8.4 +/− 0.2) Ethoxylated Macadamia Nut Oil 20 mls Ethyl AlcoholAnhydrous 20 mls

Admix in the listed order and pour off into 60 ml roll on bottles andapply directly to affected areas morning and evening.

Other formulations of transdermal delivery composition can be used torelax the muscles of a subject. By one approach, a transdermal deliverycomposition that comprises a compound that relaxes the muscles (e.g.,chlorzoxazone or ibuprofen) is provided to a subject in need of a musclerelaxant. Accordingly methods of treating or preventing muscle sorenessare embodiments.

Other formulations of transdermal delivery composition can be used toraise the levels of a hormone in a subject in need thereof. By oneapproach, a transdermal delivery composition that comprises a hormone(e.g., any one of or combination of the hormones disclosed above orderivatives or functional analogues thereof) is provided to a subject inneed thereof. Accordingly methods of treating or preventing a hormonedeficiency or methods of increasing the level of a hormone in a subjectusing one of the transdermal delivery compositions described herein areembodiments.

Other formulations of transdermal delivery composition can be used toraise the levels of a hormone, for example, growth factor in a subjectin need thereof. By one approach, a transdermal delivery compositionthat comprises a growth factor (e.g., a growth factor contained inBioserum, which is obtainable through Atrium Biotechnologies of QuebecCity, Canada) is provided to a subject in need thereof. In otherembodiments, a transdermal delivery composition comprising a peptidethat comprises the sequence LKEKK (SEQ ID NO: 1) is provided to asubject in need of an increase in a growth factor. Accordingly methodsof treating or preventing a growth factor deficiency or methods ofincreasing the level of a growth factor in a subject using one of thetransdermal delivery compositions described herein are embodiments. Byanother approach, a transdermal delivery composition that comprisesoxytocin, vasopressin, insulin, melanocyte-stimulating hormone,corticortropin, lipotropin, thyrotropin, growth hormone, prolactin,luteinizing hormone, human chorionic gonadotropin, follicle stimulatinghormone, corticotropin-releasing factor, gonadotropin-releasing factor,prolactin-releasing factor, prolactin-inhibiting factor, growth-hormonereleasing factor, somatostatin, thyrotropin-releasing factor,calcitonin, calcitonin gene-related peptide, parathyroid hormone,glucagon-like peptide 1, glucose-dependent insulinotropic polypeptide,gastrin, secretin, cholecystokinin, motilin, vasoactive intestinalpeptide, substance P, pancreatic polypeptide, peptide tyrosine tyrosine,neuropeptide tyrosine, amphiregulin, insulin, glucagon, placentallactogen, relaxin, angiotensin II, atrial natriuretic peptide,melatonin, thyroxine, triiodothyronine, estradiol, estrone,progesterone, testosterone, cortisol, corticosterone, aldosterone,epinephrine, norepinepherine, or calctriol, is provided to a subject inneed of the same.

Other formulations of the transdermal delivery composition describedherein are used to brighten the skin, reduce age spots or skindiscolorations, reduce stretch marks, reduce spider veins, or add dyes,inks, (e.g., tattoo ink), perfumes, or fragrances to the skin of asubject. In some embodiments, for example, transdermal deliverycompositions that comprise a compound that brightens the skin or reducesage spots or skin discolorations (e.g., Melaslow, a citrus-based melanin(tyrosinase) inhibitor obtainable from Revivre Laboratories of Singaporeor Etioline, a skin brightener made from an extract from the Mitracarpeleaf obtainable from Krobell, USA), or a compound that reduces stretchmarks (Kayuuputih Eucalyptus Oil, obtainable from Striad Laboratories)or add dyes, inks, (e.g., tattoo ink), perfumes, or fragrances areprovided to the skin of a subject.

It has also been discovered that ethoxylated oil by itself, preferablymacadamia nut oil having 10-19 ethoxylations/molecule (i.e., 10, 11, 12,13, 14, 15, 16, 17, 18, or 19 ethoxylations/molecule), has therapeuticand cosmetic properties. For example, application of an ethoxylated oil(e.g., macadamia nut oil having 16 ethoxylations/molecule) was found toreduce stretch marks and spider veins on a subject in need thereof.Application of an ethoxylated oil (e.g., macadamia nut oil having 16ethoxylations/molecule) to a burn (e.g., a sun burn or a skin burnobtained from over-heated metal) was found to significantly expediterecovery from the burn, oftentimes without blistering. Accordingly, someembodiments concern a transdermal delivery composition comprising anethoxylated oil (e.g., macadamia nut oil that was ethoxylated 10-19ethoxylations per molecule, 11-19 per molecule, 12-18 ethoxylations permolecule, 13-17 ethoxylations per molecule, 14-16 ethoxylations permolecule, or 15 ethoxylations per molecule) and these compositions areused to reduce the appearance of stretch marks and spider veins orfacilitate the recovery from burns of the skin.

In addition to the delivery of low and medium molecular weight deliveredagents, several compositions that have high molecular weight deliveredagents (e.g., collagens) and methods of use of such compositions areembodiments of the invention. Preferred formulations of the transdermaldelivery composition comprise a collagen (natural or synthetic) orfragment thereof at least 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 24, 30,40, 50, 100, 250, 500, 1000, 1500, 2000, 2500, 3000, 5000, or more aminoacids in length and these compositions are used to reduce wrinkles andfine lines on a subject.

For example, some embodiments concern a transdermal delivery compositioncomprising an ethoxylated fatty moiety, an ethoxylated lipid (e.g.,macadamia nut oil that was ethoxylated 10-19 ethoxylations per molecule,11-19 per molecule, 12-18 ethoxylations per molecule, 13-17ethoxylations per molecule, 14-16 ethoxylations per molecule, or 15ethoxylations per molecule), or an ethoxylated transdermal deliveryenhancer and a therapeutically effective amount of a collagen orfragment thereof (e.g., marine collagen). In some aspects of theinvention, a transdermal delivery composition comprising an ethoxylatedoil and collagen also contains water and/or an alcohol and/or an aqueousadjuvant such as Aloe Vera.

In different embodiments of this transdermal delivery composition, thecollagen has a molecular weight less than, or equal to 6,000 daltons orgreater than 6,000 daltons. Thus, in some embodiments, the collagen canhave an approximate molecular weight as low as 2,000 daltons or lower.In other embodiments, the molecular weight is from about 300,000 daltonsto about 500,000 daltons. Further, these transdermal deliverycompositions can have a therapeutically effective amount of collagen orfragment thereof by weight or volume that is 0.1% to 85.0%. The collagencan be any natural or synthetic collagen, for example, Hydrocoll EN-55,bovine collagen, human collagen, a collagen derivative, marine collagen,Solu-Coll, or Plantsol, recombinant or otherwise man made collagens orderivatives or modified versions thereof (e.g., protease resistantcollagens). As above, an apparatus having a vessel joined to anapplicator that houses the transdermal delivery composition containingcollagen is also an embodiment and preferred applicators or dispensersinclude a roll-on or a sprayer.

Accordingly, some of the embodied methods concern the reduction ofwrinkles and or the improvement of skin tone by using a transdermaldelivery composition comprising an ethoxylated oil and a collagen and/ora fragment thereof. Some formulations to be used to reduce wrinkles andimprove skin tone include an ethoxylated fatty moiety, an ethoxylatedlipid moiety (e.g., macadamia nut oil that was ethoxylated 10-19ethoxylations per molecule, 11-19 per molecule, 12-18 ethoxylations permolecule, 13-17 ethoxylations per molecule, 14-16 ethoxylations permolecule, or 15 ethoxylations per molecule), or an ethoxylatedtransdermal delivery enhancer, and a therapeutically effective amount ofa collagen or fragment thereof (e.g., marine collagen) that is at least2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 24, 30, or 40 amino acids in length.Some formulations that can be used to practice the method above includea transdermal delivery composition comprising an ethoxylated oil andcollagen or fragment thereof, as described above, and, optionally, waterand/or an alcohol and/or an aqueous adjuvant such as Aloe Vera. Forexample, by one approach, a method of reducing wrinkles or improvingskin tone is practiced by identifying a subject in need thereof andproviding said subject a transdermal delivery composition, as describedherein and, optionally, monitoring the subject for restoration orimprovement of skin tone and the reduction of wrinkles.

BRIEF DESCRIPTION OF THE DRAWINGS

Embodiments of a transdermal delivery device are depicted in FIGS. 1Athrough 4B. In aspects of the invention, a transdermal delivery systemcomprises a transdermal delivery composition and a transdermal deliverydevice.

FIG. 1A schematically depicts in an exploded state a dispenser fordelivery of a transdermal drug delivery system fluid comprising aremovable cartridge.

FIG. 1B schematically depicts the dispenser of FIG. 1A in an assembledstate.

FIG. 2 schematically depicts a cross section of the dispenser of FIG.1B.

FIG. 3A schematically depicts a cross section of the upper portion of apartially filled dosing chamber having an upper wall configured allowair to escape, but prevent fluid from escaping.

FIG. 3B schematically depicts the upper portion of the dosing chamber ofFIG. 3A, where the dosing chamber is full and fluid is prevented fromescaping.

FIG. 4A schematically depicts a cross-section of the dispenser of FIG.2, taken along the line 4, wherein the slidable member is in a firstposition permitting filling of the dosing chamber.

FIG. 4B schematically depicts the cross-section of FIG. 4A, wherein theslidable member is in a second position permitting delivery of the dosedfluid.

DETAILED DESCRIPTION OF THE INVENTION

Several transdermal delivery compositions and devices for providing saidcompositions to a subject are described herein. Embodiments of theinvention can be used to transdermally deliver low or high (or both lowand high) molecular weight pharmaceuticals, prophylactics, diagnostics,and cosmetic agents to a subject. The transdermal delivery compositionsdisclosed herein are useful for the delivery of various types ofcompounds including but not limited to nucleic acids, peptides, modifiedpeptides, small molecules, immunogenic preparations, and the like. Someembodiments include transdermal delivery compositions that canadminister compounds having molecular weights greater than 6,000daltons. One embodiment, for example, includes a transdermal deliverycomposition that can administer a therapeutically effective amount of anon-steroidal anti-inflammatory drug (NSAID). Still more embodimentsconcern transdermal delivery compositions that can administer hormones,anesthetics, collagen preparations e.g., soluble collagens, hydrolyzedcollagens, and fragments of collagen), cardiovascular pharmaceuticalcompounds, anti-infective compounds (e.g. antibiotics and antiviralcompounds), diabetes-related treatments, immunogenic compositions,vaccines, immune response modifiers, enzyme inhibitors, analgesics(e.g., a formulation comprising capsaicin or Boswellin or both),migraine therapies, sedatives, imaging and contrast compounds. Theseexamples are provided to demonstrate that embodiments of the inventioncan be used to transdermally deliver both low and high molecular weightcompounds and it should be understood that many other molecules can beeffectively delivered to the body, using the embodiments describedherein, in amounts that are therapeutically, prophylactically, orcosmetically beneficial.

Some transdermal delivery compositions described herein comprise aliposphere that is configured to deliver a wide variety of deliveredagents. As used herein, the term “liposphere” refers to a spherical orovoid-shaped structure comprising an ethoxylated or propoxylated fattymoiety, which contains or is associated with (e.g., joined to) adelivered agent. That is, the term “lipospheres” includes, but is notlimited to, liposomes that comprise an ethoxylated or propoxylated oil,fatty acid, fatty amine, or fatty alcohol. Accordingly, the term “fattymoiety” can refer to a fatty acid, a fatty alcohol, fatty amine, orother fatty acid derivative. The ethoxylated fatty moiety or lipidmoiety has both hydrophobic and hydrophilic properties, in that thehydrocarbon chain of the fatty moiety or lipid moiety is hydrophobic,and the polyethoxy groups confer hydrophilicity on the molecule. Thepreparation of propoxylated fatty moieties and lipid moieties is wellknown. (See, e.g., Raths et al., supra). Due to their similarity instructure, propoxylated fatty moieties and lipids will share many of thesame characteristics as ethoxylated fatty moieties and lipids.Accordingly, fatty moieties and lipid moieties that are propoxylated orethoxylated and propoxylated are contemplated penetration enhancers andtransdermal delivery enhancers.

In the embodiments disclosed herein, the number of ethoxylations isadjusted to between 10 and 19 ethoxylations per molecule to achieveoptimal transdermal delivery of the delivered agent. Ethoxylated fattyacids, fatty alcohols, and fatty amines, are commercially available(e.g., Ethox Chemicals, LLC, Greenville, S.C.; A&E Connock, Ltd.,Hampshire, England; Floratech, Glibert, Ariz.). Alternativelyethoxylated fatty moieties are synthesized using methods known to thoseskilled in the art (See, U.S. Pat. No. 6,300,508 to Raths et al.; U.S.Pat. No. 5,936,107 to Raths et al.) by reacting fatty moieties withethylene oxide.

By way of example, ethoxylated oils can be prepared using a two-stepprocess that starts with trans-esterification with added glycerolfollowed by ethoxylation of the product of this reaction.Trans-esterification is performed by any method available to thoseskilled in the art, such as heating an ester, such as the glycerolesters present in natural vegetable oils, in the presence of anotheralcohol or polyol, such as glycerol, in the presence of a catalyst.Catalysts useful in the transesterification reaction include gaseouscatalysts, such hydrochloric acid bubbled through the reaction mixture.Alternatively, solid catalysts such as zinc oxide or the acetates ofcopper, cobalt or zinc can also be used. The transesterificaitonreaction produces one or two fatty acids attached to a molecule ofglycerol. The ratio of mono- and di-esters can be controlled by theamount of glycerol used in the reaction (i.e. higher ratios ofglycerol:oil will yield more reactive —OH and fewer fatty acid moietiesper molecule, and a lower ratio of glycerol:oil would give more fattyacids, as is apparent to those skilled in the art. The hydroxyl groupsare subsequently reacted with ethylene oxide in the presence of anappropriate catalyst, (e.g., aluminum) using methods known to thoseskilled in the art.

Purified fatty moieties commercially available from a variety of sources(e.g., SIGMA-Aldrich, St. Louis, Mo.) are suitable for use in thetransdermal delivery compositions described above.

Alternative embodiments of transdermal delivery compositions describedherein comprise a penetration enhancer that includes an ethoxylatedlipid moiety. It was discovered that ethoxylated lipids (e.g.,ethoxylated oils) can be used as transdermal penetration enhancers inthat they effectively transport low and high molecular weight compoundsthrough the skin. It was also discovered that ethoxylated oils, bythemselves, have therapeutic and cosmetic applications (e.g., thereduction of the appearance of spider veins and stretch marks orpromoting expedited recovery from burns to the skin). Ethoxylated lipidscan be created in many ways, however, a preferred approach involves thereaction of ethylene oxide with a vegetable, nut (e.g., macadamia nut),animal, or synthetic oil. In embodiments where the transdermal deliverycomposition comprises an ethoxylated oil, it is contemplated that insome embodiments, ethoxylated fatty moieties are used to fortify orsupplement ethoxylated oils in some embodiments. By way of example,ethoxylated macadamia nut oil can be fortified with ethoxylated palmiticor oleic acid.

Several transdermal delivery enhancers disclosed herein are compoundshaving a multifunctional backbone. The multifunctional backbone can beone of many chemical structures that have at least two reactive hydrogenresidues, such that the multifunctional backbone is the basis of atransdermal delivery enhancer with least one fatty moiety and at leastone polyethoxy group. The reactive hydrogen residues (R) are present in—OH, COOH, SH, and NH₂, groups.

The polyethoxy group has the structure:

—O—(CH₂—CH₂-O—)_(n)H

Embodiments wherein n is between 10 and 19 per molecule of transdermaldelivery enhancer to possess superior transdermal delivery properties.

In preferred embodiments, the fatty moiety component of the transdermaldelivery enhancer has a carbon chain of at least 10 carbon residues. Thechain length of the fatty moiety can be for example 10, 12, 14, 16, 18,20, 22, or 24 residues. Further, the fatty moiety may be saturated,unsaturated, or polyunsaturated.

Desirably, the multifunctional backbone has at least three reactivegroups. The reactive groups can be homogeneous. For example, in someembodiments, the multifunctional backbone is a tri-alcohol comprisingthree —OH groups, such as 1, 2, 3-butanetriol, 1, 2, 4 butantetriol,pyrogallol (1,2,3-benezentriol), hydroxyquinol (1,2,4-benzenetriol),trimethyololpropane, 1, 2, 6-hexanetriol and the like. Other examples ofmultifunctional backbones suitable as the foundation of a transdermaldelivery enhancer include tri-acids, comprising three carboxylategroups, such as hemi-mellitic acid, trimellitic acid, trimesic acid,nitrilotriacetic acid, and the like. Those skilled in the art willappreciate that other tricarboxylic acids are suitable asmultifunctional backbones.

Alternative multifunctional backbones have heterogeneous reactivegroups, e.g., a combination of at least two different reactive groups(e.g., a COOH group and an NH₂ group). For example, amino acids such asglutamic acid, aspartic acid, cysteine, glutamine, serine, threonine,tryrosine, and lysine have three reactive groups and are suitable asmultifunctional backbones. Similarly, di- and tri-peptides will havethree or more reactive groups and are thus suitable as multifunctionalbackbones.

Triethanolamine, diethanolamine, dimethylolurea, and glucosamine areother exemplary multifunctional backbones with heterogeneous reactivegroups.

Simple carbohydrates are small straight-chain aldehydes and ketones withseveral hydroxyl groups, usually one on each carbon except thefunctional group. Due to the presence of the multiple —OH groups oncarbohydrates such as tetroses, pentoses, hexoses, and so forth, thesecompounds are another source of multifunctional backbones useful ascomponents of transdermal delivery enhancers. Exemplary carbohydratesthat are useful components of transdermal delivery enhancers includeglucose, mannose, fructose, ribose, xylose, threose, erythrose, and thelike.

Sugar alcohols such as sorbitol, mannitol, xylitol, erythritol,petaerythritol, and inositol are useful components of transdermaldelivery enhancers.

Not wanting to be tied to any particular mechanism or mode of action andoffered only to expand the knowledge in the field, it is contemplatedthat the ethoxylated fatty moiety, ethoxylated lipid moiety, orethoxylated multifunctional backbone encapsulates the delivered agent ina sphere-like composition, forming a “liposphere” that exhibits greatlyenhanced transdermal delivery properties.

Each of the disclosed transdermal delivery compositions can containadditional compounds such as alcohols, nonionic solubilizers oremulsifiers. In some compositions, these compounds are added to improvethe solubility of the delivered agent or effectiveness or fluidity ofthe liposphere, penetration enhancer, or transdermal delivery enhancer.Suitable hydrophilic components include, but are not limited to,ethylene glycol, propylene glycol, dimethyl sulfoxide (DMSO), dimethylpolysiloxane (DMPX), oleic acid, caprylic acid, isopropyl alcohol,1-octanol, ethanol (denatured or anhydrous), and other pharmaceuticalgrade or absolute alcohols.

Other embodiments of the transdermal delivery compositions comprise anaqueous adjuvant. Aqueous adjuvants include, but are not limited to,water (distilled, deionized, filtered, or otherwise prepared), Aloe Verajuice, and other plant extracts such as chlorophyll or Spirulina. Thus,several embodiments of the invention have a hydrophobic/hydrophiliccomponent comprising an ethoxylated fatty moiety (e.g., palmitoleicacid, oleic acid, or palmitic acid) or an ethoxylated oil (e.g.,macadamia nut oil, coconut oil, eucalyptus oil, synthetic oils, castoroil, glycerol, corn oil, jojoba oil, or emu oil) and may contain ahydrophilic component comprising an alcohol, a nonionic solubilizer, oran emulsifier (e.g., isopropyl alcohol) and/or, optionally, an aqueousadjuvant, such as water and/or Aloe Vera extract.

Other materials can also be components of a transdermal deliverycomposition of the invention including fragrance, creams, ointments,colorings, and other compounds so long as the added component does notdeleteriously affect transdermal delivery of the delivered agent. It hasbeen found that the Aloe Vera, which allows for transdermal delivery ofhigh molecular weight delivered agents, including collagen having anaverage molecular weight greater than 6,000 daltons, can be removed fromtransdermal delivery compositions comprising a light oil (e.g.,macadamia nut oil) that has been ethoxylated to the range of 10-19ethoxylations/molecule. Formulations lacking Aloe Vera provide theunexpected benefit of efficient transdermal delivery, uniformapplication and quick penetration making these formulations superior toformulations that contain Aloe Vera.

Similarly, formulations of transdermal delivery compositions that lackalcohol provide the unexpected benefit of efficient transdermaldelivery, uniform application, and quick penetration without the dryingor irritation brought about by the alcohol. Additionally, formulationslacking water or other aqueous adjuvants provide efficient transdermaldelivery while maintaining the highest possible concentration ofdelivered agent and, also, provide for quick penetration without theskin-drying effects seen with some formulations that contain alcohol.

A molecule or a mixture of molecules (e.g., a pharmaceutical, chemical,or cosmetic agent) that are delivered to the body using an embodiment ofa transdermal delivery composition are termed “delivered agents”. Adelivered agent that can be administered to the body using an embodimentof the invention can include, for example, a protein or peptide, asugar, a nucleic acid, a chemical, a lipid, or derivatives of the same.Desirable delivered agents include, but are not limited to,glycoproteins, enzymes, genes, nucleic acids, peptides, drugs, andceramides. Preferred delivered agents include NSAIDS, collagens orfragments thereof, capsaicin, and Boswellin. In some embodiments, atransdermal delivery composition comprises a combination of any two ofthe aforementioned delivered agents. Other delivered agents include, forexample, hormones, anti-inflammatory drugs, anesthetics, analgesics,sedatives, migraine therapies, cardiovascular pharmaceuticals,anti-infective agents, diabetes-related therapies, vaccines, imagingagents, contrast agents, glucosamine, chondroitin sulfate, MSM,perfumes, melasin, nicotine, nicotine analogs, peptides, amino acids,nucleic acids, and peptidomimetics.

In addition to the aforementioned compositions, methods of making andusing the embodiments of the invention are provided. In one aspect, atransdermal delivery composition is prepared by mixing an ethoxylatedfatty moiety with a delivered agent.

In another aspect, a transdermal delivery composition is prepared bymixing a hydrophilic component with a hydrophobic component and anaqueous adjuvant. Depending on the solubility of the delivered agent,the delivered agent can be solubilized in either the ethoxylated oil, ahydrophobic, hydrophilic, or aqueous adjuvant or water prior to mixing.

In addition to physical mixing techniques, (e.g., magnetic stirring orrocker stirring), embodiments of the methods contemplate heat can beapplied to help coalesce the mixture. Desirably, the temperature is notraised above 40° C.

Several formulations of transdermal delivery compositions are within thescope of aspects of the invention. In embodiments wherein thetransdermal delivery composition includes an aqueous adjuvant, infurther embodiments, the formulation comprises a ratio of hydrophiliccomponent:hydrophobic component:aqueous adjuvant of 3:4:3. The amount ofdelivered agent that is incorporated into the penetration enhancerdepends on the compound, desired dosage, and application. The amount ofdelivered agent in a particular formulation can be expressed in terms ofpercentage by weight, percentage by volume, or concentration. Severalspecific formulations of delivery systems are provided in the Examplesdescribed herein.

Methods of treatment and prevention of pain, inflammation, and humandisease are also provided. In some embodiments, a transdermal deliverycomposition comprising an NSAID, capsaicin, Boswellin or any combinationthereof is provided to a patient in need of treatment, such as forrelief of pain and/or inflammation. A patient can be contacted with thetransdermal delivery composition and treatment continued for a timesufficient to reduce pain or inflammation or inhibit the progress ofdisease.

Additionally, a method of reducing wrinkles, removing age spots, andincreasing skin tightness and flexibility is provided. By this approach,a transdermal delivery composition comprising a collagen or fragmentthereof or melaslow or other skin brightening agent is provided to apatient in need, the patient is contacted with the transdermal deliverycomposition, and treatment is continued for a time sufficient to restorea desired skin tone (e.g., reduce wrinkles, age spots, or restore skinbrightness, tightness and flexibility). In the disclosure below, thereis provided a description of several of the delivered agents that can beincorporated into the transdermal delivery compositions describedherein.

Delivered Agents

Many different delivered agents can be incorporated into the varioustransdermal delivery compositions described herein. While thetransdermal delivery of molecules having a molecular weight in thevicinity of 6000 daltons has been reported, it has not been possible,until the present invention, to administer molecules of greater sizetransdermally. (See U.S. Pat. No. 5,614,212 to D'Angelo et al.).

The described embodiments can be organized according to their ability todeliver a low or high molecular weight delivered agent. Low molecularweight molecules (e.g., a molecule having a molecular weight less than6,000 daltons) can be effectively delivered using an embodiment of theinvention and high molecular weight molecules (e.g., a molecule having amolecular weight greater than 6,000 daltons) can be effectivelydelivered using an embodiment of the invention. Desirably, a transdermaldelivery composition described herein provides a therapeutically,prophylactically, diagnostically, or cosmetically beneficial amount of adelivered agent having a molecular weight of 50 daltons to less than6,000 daltons. Preferably, however, a transdermal delivery compositiondescribed herein provides a therapeutically, prophylactically,diagnostically, or cosmetically beneficial amount of a delivered agenthaving a molecular weight of 50 daltons to 2,000,000 daltons or less.That is, a transdermal delivery composition described herein,preferably, provides a delivered agent having a molecular weight of lessthan or equal to or greater than 50, 100, 200, 500, 1,000, 1,500, 2,000,2,500, 3,000, 3,500, 4,000, 4,500, 5,000, 5,500, 6,000, 7,000, 8,000,9,000, 10,000, 11,000, 12,000, 13,000, 14,000, 15,000, 16,000, 17,000,18,000, 19,000, 20,000, 21,000, 22,000, 23,000, 24,000, 25,000, 26,000,27,000, 28,000, 29,000, 30,000, 31,000, 32,000, 33,000, 34,000, 35,000,36,000, 37,000, 38,000, 39,000, 40,000, 41,000, 42,000, 43,000, 44,000,45,000, 46,000, 47,000, 48,000, 49,000, 50,000, 51,000, 52,000, 53,000,54,000, 55,000, 56,000, 57,000, 58,000, 59,000, 60,000, 61,000, 62,000,63,000, 64,000, 65,000, 66,000, 67,000, 68,000, 69,000, 70,000, 75,000,80,000, 85,000, 90,000, 95,000, 100,000, 125,000, 150,000, 175,000,200,000, 225,000, 250,000, 275,000, 300,000, 350,000, 400,000, 450,000,500,000, 600,000, 700,000, 800,000, 900,000, 1,000,000, 1,500,000,1,750,000, and 2,000,000 daltons.

In one aspect, a low molecular weight compound (e.g., a pain relievingsubstance or mixture of pain relieving substances) is transdermallydelivered to cells of the body using an embodiment described herein. Thedelivered agent can be, for example, any one or more of a number ofcompounds, including non-steroidal anti-inflammatory drugs (NSAIDs) thatare frequently administered systemically. These include ibuprofen(2-(isobutylphenyl)-propionic acid); methotrexate (N-[4-(2, 4 diamino6-pteridinyl-methyl]methylamino]benzoyl)-L-glutamic acid); aspirin(acetylsalicylic acid); salicylic acid; diphenhydramine(2-(diphenylmethoxy)-N,N-dimethylethylamine hydrochloride); naproxen(2-naphthaleneacetic acid, 6-methoxy-9-methyl-, sodium salt, (−));phenylbutazone (4-butyl-1,2-diphenyl-3,5-pyrazolidinedione);sulindac-(2)-5-fluoro-2-methyl-1-[[p-(methylsulfinyl)phenyl]methylene-]-1H-indene-3-aceticacid; diflunisal (2′,4′,-difluoro-4-hydroxy-3-biphenylcarboxylic acid;piroxicam (4-hydroxy-2-methyl-N-2-pyridinyl-2H-1,2-benzothiazine-2-carboxamide 1,1-dioxide, an oxicam; indomethacin(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-H-indole-3-acetic acid);meclofenamate sodium (N-(2,6-dichloro-m-tolyl) anthranilic acid, sodiumsalt, monohydrate); ketoprofen (2-(3-benzoylphenyl)-propionic acid;tolmetin sodium (sodium 1-methyl-5-(4-methylbenzoyl-1H-pyrrole-2-acetatedihydrate); diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneaticacid, monosodium salt); hydroxychloroquine sulphate(2-{[4-[(7-chloro-4-quinolyl)amino]pentyl]ethylamino}ethanol sulfate(1:1); penicillamine (3-mercapto-D-valine); flurbiprofen([1,1-b]phenyl]-4-acetic acid, 2-fluoro-alphamethyl-, (+−.)); cetodolac(1-8-diethyl-13,4,9, tetrahydropyrano-[3-4-13]indole-1-acetic acid;mefenamic acid (N-(2,3-xylyl)anthranilic acid; and diphenhydraminehydrochloride (2-diphenyl methoxy-N, N-di-methylethamine hydrochloride).

The transdermal delivery compositions described herein, which containNSAIDs, desirably comprise an amount of the compound that istherapeutically beneficial for the treatment or prevention of disease orinflammation. Several studies have determined an appropriate dose of anNSAID for a given treatment or condition. (See e.g., Woodin, RN, August:26-33 (1993) and Amadio et al., Postgrduate Medicine, 93(4):73-97(1993)). The maximum recommended daily dose for several NSAIDs is listedin TABLE 1.

A sufficient amount of NSAID can be incorporated into a transdermaldelivery composition described herein such that a therapeuticallyeffective amount of NSAID is effectively delivered to a subject. Forexample, about 0.5 ml of the transdermal delivery composition describedherein is applied in a single application. A therapeutically effectiveamount of ibuprofen is about 800 mg/dose. Accordingly, a 30 ml bottlecontaining a transdermal delivery system formulation and ibuprofen cancontain 24 grams of ibuprofen such that 800 mg of ibuprofen is providedin each 1.0 ml. Because the transdermal delivery compositions describedherein can provide a delivered agent in a site-specific manner, a lowertotal dose of therapeutic agent, as compared to the amounts providedsystemically, will provide therapeutic benefit. Additionally, greatertherapeutic benefit can be gained by using a transdermal deliverycomposition described herein because a greater concentration oftherapeutic agent (e.g., an NSAID) can be provided to the particularsite of inflammation. That is, in contrast to systemic administration,which applies the same concentration of therapeutic to all regions ofthe body, a transdermal delivery composition can site-specificallyprovide the therapeutic agent and, thereby, provide a much greaterregional concentration of the agent than if the same amount oftherapeutic were administered systemically.

TABLE 1 Agent Maximum Recommended Daily Dose Indomethacin 100 mgIbuprofen 3200 mg  Naproxen 1250 mg  Fenoprofen 3200 mg  Tolmetin 2000mg  Sulindac 400 mg Meclofenamate 400 mg Ketoprofen 300 mg Proxicam  10mg Flurbiprofen 300 mg Diclofenac 200 mg

Additional embodiments include a transdermal delivery composition thatprovides a pain relieving mixture comprising capsaicin (e.g., oleoresincapsicum) or Boswellin or both. Capsaicin(8-methyl-N-vanillyl-6-nonenamide), the pungent component of paprika andpeppers, is a potent analgesic. (See U.S. Pat. Nos. 5,318,960 to Toppo,5,885,597 to Botknecht et al., and 5,665,378 to Davis et al., hereinexpressly incorporated by reference in their entireties). Capsaicinproduces a level of analgesia comparable to morphine, yet it is notantagonized by classical narcotic antagonists such as naloxone. Further,it effectively prevents the development of cutaneous hyperalgesia, butappears to have minimal effects on normal pain responses at moderatedoses. At high doses capsaicin also exerts analgesic activity inclassical models of deep pain, elevating the pain threshold above thenormal value. Capsaicin can be readily obtained by the ethanolextraction of the fruit of Capsicum frutescens or Capsicum annum.Capsaicin and analogs of capsaicin are available commercially from avariety of suppliers, and can also be prepared synthetically bypublished methods. Aspects of the invention encompass the use ofsynthetic and natural capsaicin, capsaicin derivatives, and capsaicinanalogs.

A form of capsaicin used in several desirable embodiments is oleoresincapsicum. Oleoresin capsicum contains primarily capsaicin,dihydrocapsaicin, nordihydrocapsaicin, homocapsaicin, andhomodihydrocapsaicin. The term “capsaicin” collectively refers to allforms of capsaicin, capsicum, and derivatives or modifications thereof.The pungency of these five compounds, expressed in Scoville units, isprovided in TABLE 2.

TABLE 2 Compound Pungency × 100,000 SU Capsaicin 160 Dihydrocapsaicin160 Nordihydrocapsaicin 91 Homocapsaicin 86 Homodihydrocapsaicin 86

The transdermal delivery compositions that are formulated to containcapsaicin desirably comprise by weight or volume 0.01% to 1.0% capsaicinor 1.0% to 10% oleoresin capsicum. Preferred amounts of this deliveredagent include by weight or volume 0.02% to 0.75% capsaicin or 2.0% to7.0% oleoresin capsicum. For example, the transdermal deliverycompositions that contain capsaicin can comprise by weight or volumeless than or equal to 0.01%, 0.015%, 0.02%, 0.025%, 0.03%, 0.035%,0.04%, 0.045%, 0.05%, 0.055%, 0.06%, 0.065%, 0.07%, 0.075%, 0.08%,0.085%, 0.09%, 0.095%, 0.1%, 0.15%, 0.175%, 0.2%, 0.225%, 0.25%, 0.275%,0.3%, 0.325%, 0.35%, 0.375%, 0.4%, 0.425%, 0.45%, 0.475%, 0.5%, 0.55%,0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, and 1.0% capsaicin.The transdermal delivery compositions of that contain capsaicin can alsocomprise an amount of capsaicin by weight or volume that is greater than1.0%, such as 1.2%, 1.5%, 1.8%, 2.0%, 2.2%, 2.5%, 2.8%, 3.0%, 3.5%,4.0%, 4.5%, and 5.0%. Similarly, the transdermal delivery compositionsthat contain oleoresin capsicum can comprise an amount of oleoresincapsicum less than 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%,5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0%, 11.0%,12.0%, and 13.0%.

Boswellin, also known as Frankincense, is an herbal extract of a tree ofthe Boswellia family. Boswellin can be obtained, for example, fromBoswellia thurifera, Boswellia carteri, Boswellia sacra, and Boswelliaserrata. There are many ways to extract Boswellin and Boswellin gumresin and boswellic acids are obtainable from several commercialsuppliers (a 65% solution of Boswellic acid is obtainable from Nature'sPlus). Some suppliers also provide creams and pills having Boswellinwith and without capsaicin and other ingredients. Embodiments of theinvention comprise Boswellin and the term “Boswellin” collectivelyrefers to Frankincense, an extract from one or more members of theBoswellia family, Boswellic acid, synthetic Boswellin, or modified orderivatized Boswellin.

The transdermal delivery compositions that contain Boswellin desirablycomprise 0.1% to 10% Boswellin by weight or volume. Preferred amounts ofthis delivered agent include 1.0% to 5.0% Boswellin by weight. Forexample, the transdermal delivery compositions that contain Boswellincan comprise by weight or volume less than or equal to 0.1%, 0.15%,0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%,0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%, 1.1%, 1.15%, 1.2%, 1.25%, 1.3%,1.35%, 1.4%, 1.45%, 1.5%, 1.55%, 1.6%, 1.65%, 1.7%, 1.75%, 1.8%, 1.85%,1.9%, 1.95%, and 2.0%, 2.1%, 2.15%, 2.2%, 2.25%, 2.3%, 2.35%, 2.4%,2.45%, 2.5%, 2.55%, 2.6%, 2.65%, 2.7%, 2.75%, 2.8%, 2.85%, 2.9%, 2.95%,3.0%, 3.1%, 3.15%, 3.2%, 3.25%, 3.3%, 3.35%, 3.4%, 3.45%, 3.5%, 3.55%,3.6%, 3.65%, 3.7%, 3.75%, 3.8%, 3.85%, 3.9%, 3.95%, 4.0%, 4.1%, 4.15%,4.2%, 4.25%, 4.3%, 4.35%, 4.4%, 4.45%, 4.4%, 4.45%, 4.5%, 4.55%, 4.6%,4.65%, 4.7%, 4.75%, 4.8%, 4.85%, 4.9%, 4.95%, and 5.0% Boswellin. Thetransdermal delivery compositions that contain Boswellin can alsocomprise amounts of Boswellin by weight that are greater than 5.0%, suchas 5.5%, 5.7%, 6.0%, 6.5%%, 6.7%, 7.0%, 7.5%, 7.7%, 8.0%, 8.5%, 8.7%,9.0%, 9.5%, 9.7%, and 10.0% or greater. Additionally, Boswellin fromdifferent sources can be combined to compose the Boswellin component ofan embodiment. For example, in one embodiment an extract from Boswelliathurifera is combined with an extract from Boswellia serrata.

Additional embodiments of the invention comprise a transdermal deliverycomposition that can administer a pain relieving solution comprising twoor more members selected from the group consisting of NSAIDs, capsacin,and Boswellin. The transdermal delivery compositions that include two ormore members selected from the group consisting of NSAIDs, capsacin, andBoswellin desirably comprise an amount of delivered agent that can beincluded in a delivered agent having an NSAID, capsaicin, or Boswellinby itself. For example, if the delivered agent comprises an NSAID, theamount of NSAID that can be used can be an amount recommended in theliterature (See e.g., Woodin, RN, August: 26-33 (1993) and Amadio, etal., Postgrduate Medicine, 93(4):73-97 (1993)), or an amount listed inTABLE 1. Similarly, if capsaicin is a component of the delivered agentsthen the transdermal delivery composition can comprise by weight orvolume less than or equal to 0.01%, 0.015%, 0.02%, 0.025%, 0.03%,0.035%, 0.04%, 0.045%, 0.05%, 0.055%, 0.06%, 0.065%, 0.07%, 0.075%,0.08%, 0.085%, 0.09%, 0.095%, 0.1%, 0.15%, 0.175%, 0.2%, 0.225%, 0.25%,0.275%, 0.3%, 0.325%, 0.35%, 0.375%, 0.4%, 0.425%, 0.45%, 0.475%, 0.5%,0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, and 1.0%capsaicin or less than 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%,5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5%, 10.0%,11.0%, 12.0%, 13.0%, oleoresin capsicum. Further, if Boswellin is acomponent of the delivered agents, then the delivery system can compriseby weight or volume less than or equal to 0.1%, 0.15%, 0.2%, 0.25%,0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%,0.85%, 0.9%, 0.95%, 1.0%, 1.1%, 1.15%, 1.2%, 1.25%, 1.3%, 1.35%, 1.4%,1.45%, 1.5%, 1.55%, 1.6%, 1.65%, 1.7%, 1.75%, 1.8%, 1.85%, 1.9%, 1.95%,2.0%, 2.1%, 2.15%, 2.2%, 2.25%, 2.3%, 2.35%, 2.4%, 2.45%, 2.5%, 2.55%,2.6%, 2.65%, 2.7%, 2.75%, 2.8%, 2.85%, 2.9%, 2.95%, 3.0%, 3.1%, 3.15%,3.2%, 3.25%, 3.3%, 3.35%, 3.4%, 3.45%, 3.5%, 3.55%, 3.6%, 3.65%, 3.7%,3.75%, 3.8%, 3.85%, 3.9%, 3.95%, 4.0%, 4.1%, 4.15%, 4.2%, 4.25%, 4.3%,4.35%, 4.4%, 4.45%, 4.4%, 4.45%, 4.5%, 4.55%, 4.6%, 4.65%, 4.7%, 4.75%,4.8%, 4.85%, 4.9%, 4.95%, 5.0%, 5.5%, 5.7%, 6.0%, 6.5%%, 6.7%, 7.0%,7.5%, 7.7%, 8.0%, 8.5%, 8.7%, 9.0%, 9.5%, 9.7%, and 10.0% Boswellin.

Other analgesics are useful delivered agents in the transdermal deliverycompositions described herein. For example, tramadol hydrochloride,fentanyl, metamizole, morphine sulphate, ketorolac tromethamine,hydrocodone, oxycodone, morphine and loxoprofen sodium are deliveredagents in certain embodiments.

Steroidal anti-inflammatory compounds are also useful delivered agentsin the transdermal delivery compositions described herein. For example,hydrocortisone, prednisolone, triamcinolone, and priroxicam aredelivered agents in certain embodiments.

Local anesthetics are low molecular weight compounds that are useful asdelivered agents in the transdermal delivery compositions describedherein. The transdermal delivery compositions disclosed herein areparticularly useful in the context of local anesthetics, where a local,concentrated dose of a delivered agent is desirable. Embodiments of thetransdermal delivery compositions include local anesthetics, such asarticaine, procaine, tetracaine, chloroprocaine and benzocaine,novocain, mepivicaine, bupivicaine, benzocaine, and lidocaine, and thelike. The maximum single dose for local anesthetic solutions issomewhere between 70 mg to 500 mg, depending upon the age and health ofthe patient.

Compounds that have anti-infective activity are also useful in thepresent invention, particularly in the context of dermal bacterial,fungal, or viral infections. Antibiotics are compounds that either killbacterial or fungal cells, or prevent them from multiplying. Severalantibiotics are known to those skilled in the art and are deliveredagents in certain embodiments of the transdermal delivery compositions,including but not limited to amoxicillin, clavulanate potassium,itraconazole, acyclovir, fluconazole, terbinafine hydrochloride,erythromycin ethylsuccinate, acetyl sulfisoxazole, penicillin V,cephalexin, erythromycin, azithromycin, tetracycline, ciproflaxin,gentamycin, sulfathiazole, nitrofurantoin, norfloxacin, flumequine, andibafloxacin, metronidazole, and nystatin. Likewise, several compoundsthat have antiviral activity useful as delivered agents include but arenot limited to acyclovir, lamivudine, indinavir sulfate, and stavudine.Those skilled in the art will appreciate that analogs and derivatives ofthe anti-infective compounds now known (e.g. valacyclovir) anddiscovered in the future are contemplated in the present invention.

In addition to low molecular weight delivered agents, many mediummolecular weight delivered agents (e.g., humates) can be delivered tocells in the body by using an embodiment of the transdermal deliverycomposition. Synthetic humates (“hepsyls”) are medium molecular weightcompounds (1,000 to 100,000 daltons), which are known to be strongantiviral and antimicrobial medicaments. (See International ApplicationPublication No. WO 9834629 to Laub). Hepsyls are generally characterizedas polymeric phenolic materials comprised of conjugated aromatic systemsto which are attached hydroxyl, carboxyl, and other covalently boundfunctional groups. A transdermal delivery composition that can providehepsyls to cells of the body has several pharmaceutical uses, includingbut not limited to, treatment of topical bacterial and viral infections.

Accordingly, in another aspect of the invention, a transdermal deliverysystem that can provide a medium molecular weight compound (e.g., a formof hepsyl) to cells of the body is provided. As described above, manydifferent medium molecular weight compounds can be provided using anembodiment of a transdermal delivery composition described herein andthe use of a medium molecular weight hepsyl as a delivered agent isintended to demonstrate that embodiments of the invention can delivermany medium molecular weight compounds to cells of the body.

In some embodiments, amino acids, peptides, nucleotides, nucleosides,and nucleic acids are transdermally delivered to cells in the body usingan embodiment of the transdermal delivery composition described herein.That is, any amino acid or peptide having at least, less than, morethan, or equal to 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 75, 100,125, 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000,2500, 3000, 3500, 4000, 4500, 5000, 7000, or 10,000 amino acids can beincorporated into a transdermal delivery composition described hereinand said delivered agent can be delivered to cells in the body shortlyafter application of the composition. These embodiments can be used, forexample, to stimulate an immune response, promote wound healing, inducecollagen synthesis, or to supplement collagen. These embodiments arealso useful for the delivery of peptide hormones. Non-limiting examplesof peptide hormones that are delivered agents in certain embodimentsinclude oxytocin (SEQ ID NO:2), vasopressin (SEQ ID NO:3),melanocyte-stimulating hormone (SEQ ID NO:4 (alpha) SEQ ID NO:5 (beta)SEQ ID NO:6 (gamma)), corticortropin (SEQ ID NO:7), lipotropin (SEQ IDNO:8 (beta) SEQ ID NO:9 (gamma)), thyrotropin (SEQ iD NO: 10), growthhormone (SEQ ID NO: 1), prolactin (SEQ ID NO: 11), luteinizing hormone(SEQ ID NO:12), human chorionic gonadotropin (available fromSIGMA-Aldrich, St. Louis, Mo., Cat. No. C1063), follicle stimulatinghormone, corticotropin-releasing factor (SEQ ID NO: 13)gonadotropin-releasing factor (SEQ ID NO:43), prolactin-releasing factor(SEQ ID NO:14), prolactin-inhibiting factor (SEQ ID NO:15),growth-hormone releasing factor (SEQ ID NO: 16), somatostatin (SEQ IDNO: 17), thyrotropin-releasing factor (SEQ ID NO:18), calcitonin (SEQ IDNO: 19), calcitonin gene-related peptide (SEQ ID NO:20), parathyroidhormone (SEQ ID NO:21), glucagon-like peptide 1 (SEQ ID NO:22),glucose-dependent insulinotropic polypeptide (SEQ ID NO:23), gastrin(SEQ ID NO:24), secretin (SEQ ID NO:25), cholecystokinin (SEQ ID NO:26),motilin (SEQ ID NO:27), vasoactive intestinal peptide (SEQ ID NO:28),substance P (SEQ ID NO:30), pancreatic polypeptide (SEQ ID NO:31),peptide tyrosine tyrosine (SEQ ID NO:32), neuropeptide tyrosine (SEQ IDNO:33), amphiregulin (SEQ ID NO:34), insulin (available from SIGMAAldrich, St. Louis, Mo., Cat. No. 1643), glucagon (SEQ ID NO:35),placental lactogen (SEQ ID NO:37), relaxin (SEQ ID NO:38), inhibin A(SEQ ID NO:39), Inhibin B (SEQ ID NO:40), Endorphins (e.g., SEQ IDNO:41), angiotensin II (SEQ ID NO:42), atrial natriuretic peptide (SEQID NO:),

Several other hormones are not peptide hormones, but are neverthelesssuitable delivered agents in embodiments of the invention. Accordingly,embodiments of the invention include cortisol (available from SIGMAAldrich, St. Louis, Mo., Cat. No. H3160), corticosterone (available fromSIGMA Aldrich, St. Louis, Mo., Cat. No. C27840), aldosterone (availablefrom SIGMA Aldrich, St. Louis, Mo., Cat. No. 05521), epinephrine(available from SIGMA Aldrich, St. Louis, Mo., Cat. No. 02252),noepinephrine (available from SIGMA Aldrich, St. Louis, Mo., Cat. No.74460), calcitriol (available from SIGMA Aldrich, St. Louis, Mo., Cat.No. 17936), progesterone (available from SIGMA Aldrich, St. Louis, Mo.,Cat. No. P8783), testosterone (available from SIGMA Aldrich, St. Louis,Mo., Cat. No. T1500), androstene (available from SIGMA Aldrich, St.Louis, Mo.) and melatonin (available from SIGMA Aldrich, St. Louis, Mo.,Cat. No. 63610).

Any nucleotide or nucleoside, modified nucleotide or nucleoside, ornucleic acid having at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,75, 100, 125, 150, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1500,2000, 2500, 3000, 3500, 4000, 4500, 5000, 7000, or 10,000 or morenucleotides can be incorporated into a transdermal delivery compositiondescribed herein and said delivered agent can be delivered to cells inthe body shortly after application of the composition. These embodimentscan also be used, for example, to stimulate an immune response, promotewound healing, or induce collagen synthesis.

Several nucleic acid immunogens and/or vaccines and therapies are knownin the art and are useful as delivered agents in embodiments of thetransdermal delivery compositions disclosed herein. Several nucleic acidimmunogens that induce an immune response (both humoral and cellular)upon administration to a host have been described. DNA vaccines forseveral viruses, as well as for tumors, are known. Those skilled in theart will appreciate that nucleic acid immunogens contain essentialregulatory elements such that upon administration to a host, theimmunogen is able to direct host cellular machinery to producetranslation products encoded by the respective delivered nucleic acids.Furthermore, those skilled in the art will appreciate that the specificsequences disclosed herein are non-limiting, and that while Applicantsreference specific nucleic acids, allelic variants, fragments of nucleicacids, as well as orthologs and paralogs, now known or later discoveredsuch as those made publicly available on databases such as Genbank™ arecontemplated in the present invention.

Several immunogens for Human Immunodeficiency Virus (HIV), have beendescribed. International Publication No. WO 01/46393 teaches thatcompositions comprising the nucleic acid encoding the HIV Nef gene,fragments thereof, or variants that are optimized for efficacy asvaccines in humans, are capable of inducing a cellular immune responsein a host. The HIV Nef protein has been shown to promote viralreplication. DNA sequences comprising the Nef sequence, including thesequences of SEQ ID NOs:52, 53, and 54 are known to be capable ofinducing a cellular immune response in individuals. InternationalPublication No. WO 04/050856 discloses that DNA vaccines comprising thenucleic acid sequences and variants of HIV gp120 (SEQ ID NOs:153, 154,155, 156) and a codon-optimized nucleic acid encoding HIV-1 Gag (SEQ IDNO:152) are capable of inducing antibody and humoral immune responses.Nucleic acids encoding HIV-1 Gag and variants thereof have also beenshown to induce an immune response when administered to a host (Qui etal., 2000, J. Virology. 74(13):5997-6005). Any of the above sequencesfrom HIV are useful delivered agents for the transdermal deliverycompositions disclosed herein.

Influenza A is the causative agent of the flu in humans. Flu epidemicscause morbidity and mortality worldwide, and each year in the USA alonemore than 200,000 patients are admitted to hospitals because ofinfluenza and there are approximately 36,000 influenza-related deaths.Immunogens directed against Influenza A generally comprise attenuatedstrains of the virus. WO 04/060720 teaches that a DNA vaccine comprisingnucleic acids of sequence SEQ ID NO:51 are capable of inducing acellular immune response against Influenza virus A.

Much work has also been done on nucleic acid-based immunogens andvaccines for the hepatitis viruses, such as hepatitis C, hepatitis B andhepatitis A. (“HCV”, “HBV”, and “HAV”) The amino acid sequence encodedby the complete coding sequence of the prototype HCV-1 genome (HCVgp1)is provided (SEQ ID NO:128). Houghton et al. (U.S.S.N. 2002/0002272)disclose nucleic acids that encode several portions of HCVgp1 that arecapable of inducing a humoral immune response. For example, nucleicacids encoding the HCV E2 envelope protein or portions thereof (SEQ IDNOs: 129, 130, 131, 132), or nucleic acids encoding both HCVE1/E2envelope proteins (SEQ ID NOs:133, 134) were capable of eliciting animmune response. Schiver et al. (International Pub. No. WO 01/43693)disclose other nucleic acid sequences from HCV that elicit protectiveimmune responses, including the sequences of SEQ ID NO's:52, 53, 54.

Embodiments of the present invention also contemplate sequences fromHBV, such as nucleic acids that encode HBV core antigen (SEQ ID NO:135);HBVsAg (Genbank™ Accession No. AR141190), and the like. Additionally,nucleic acid sequences from the HAV genome (Genbank™ Accession No.NC_(—)001489) are contemplated as delivered agents.

Various other nucleic acid-based immunogens and vaccines against viralpathogens have been described in the art, such as vaccines comprisingnucleic acids from Hantavirus. Hantavirus is the causative agent ofHantavirus Pulmonary Syndrome (HPS), a form of adult respiratory diseasesyndrome that is potentially fatal in humans. WO 04/058808 disclosessequences (SEQ ID NOs:126, 127) that are useful delivered agents. Chen(International Pub. No. WO 04/110483) discloses several amino acidsequences, (SEQ ID NOs:147, 148, 149 150), the encoding nucleic acidsequences of which are useful as delivered agents for vaccines SARS.

Vaccines and immunogens comprising nucleic acids that encode a member ofthe Inhibitor of Apoptosis (IAP) family of proteins are also useful inthe context of cancer treatment. For Example, Xiang et al.(International Publication No. WO 04/099389) teach DNA vaccinescomprising sequences encoding members of the Inhibitor of Apoptosis(IAP) family of proteins, such as nucleic acids encoding the sequencesof SEQ ID NO's:136, 137, 138, and 139. These sequences are also usefulas delivered agents in one or more of the transdermal delivery systemsdescribed herein for the purposes of anti-tumor therapy.

Globulins constitute a diverse group of proteins that share the commoncharacteristic of being soluble in water or dilute salt solutions. Dueto their ability to specifically bind to target antigens, antibodies arean extremely valuable example of globulins. Offered as non-limitingexamples of therapeutically valuable antibody therapies that arecontemplated as delivered agents are: e.g. Rituxin™ for lymphoma; humanrabies immune globulin (HRIG) for rabies; bacterial polysaccharideimmune globulin (BPIG) for passive immunization of infants againstbacterial infections; pertussis immune globulin, and Herceptin™ forbreast cancer. Those skilled in the art will appreciate that a widevariety of antibody therapies-polyclonal, monoclonal, includingchemically or physically modified antibodies—can be used as deliveredagents in the transdermal delivery compositions described herein.

Other globulins are involved in the transport of a variety ofsubstances, including lipids, hormones, and inorganic ions. For example,sex-hormone binding globulin binds to and transports testosterone, andto a lesser degree estrogens, and thyronine-binding globulin binds tothyoxine. Globulins other than immunoglobulins are also contemplated asdelivered agents in the transdermal delivery system described herein.

Immune response modifiers (“IRMs”) are compounds that act on the immunesystem by inducing and/or suppressing cytokine biosynthesis. IRMspossess potent immunostimulating activity including, but not limited to,antiviral and antitumor activity, and can also down-regulate otheraspects of the immune response, for example shifting the immune responseaway from a TH₂ immune response, which is useful for treating a widerange of TH₂ mediated diseases. IRMs can also be used to modulatehumoral immunity by stimulating antibody production by B cells. SomeIRMs are small organic compounds having a molecular weight under about1000 daltons, preferably under about 500 daltons.

Examples of classes of small molecule IRM compounds include, but are notlimited to, compounds having a 2-aminopyridine fused to a five-memberednitrogen-containing heterocyclic ring. Such compounds include, forexample, imidazoquinoline amines including, but not limited to,substituted imidazoquinoline amines such as, for example, amidesubstituted imidazoquinoline amines, sulfonamide substitutedimidazoquinoline amines, urea substituted imidazoquinoline amines, arylether substituted imidazoquinoline amines, heterocyclic ethersubstituted imidazoquinoline amines, amido ether substitutedimidazoquinoline amines, sulfonamido ether substituted imidazoquinolineamines, urea substituted imidazoquinoline ethers, thioether substitutedimidazoquinoline amines, and 6-, 7-, 8-, or 9-aryl or heteroarylsubstituted imidazoquinoline amines; tetrahydroimidazoquinoline aminesincluding, but not limited to, amide substitutedtetrahydroimidazoquinoline amines, sulfonamide substitutedtetrahydroimidazoquinoline amines, urea substitutedtetrahydroimidazoquinoline amines, aryl ether substitutedtetrahydroimidazoquinoline amines, heterocyclic ether substitutedtetrahydroimidazoquinoline amines, amido ether substitutedtetrahydroimidazoquinoline amines, sulfonamido ether substitutedtetrahydroimidazoquinoline amines, urea substitutedtetrahydroimidazoquinoline ethers, and thioether substitutedtetrahydroimidazoquinoline amines; imidazopyridine amines including, butnot limited to, amide substituted imidazopyridine amines, sulfonamidesubstituted imidazopyridine amines, urea substituted imidazopyridineamines, aryl ether substituted imidazopyridine amines, heterocyclicether substituted imidazopyridine amines, amido ether substitutedimidazopyridine amines, sulfonamido ether substituted imidazopyridineamines, urea substituted imidazopyridine ethers, and thioethersubstituted imidazopyridine amines; 1,2-bridged imidazoquinoline amines;6,7-fused cycloalkylimidazopyridine amines; imidazonaphthyridine amines;tetrahydroimidazonaphthyridine amines; oxazoloquinoline amines;thiazoloquinoline amines; oxazolopyridine amines; thiazolopyridineamines; oxazolonaphthyridine amines; thiazolonaphthyridine amines; and1H-imidazo dimers fused to pyridine amines, quinoline amines,tetrahydroquinoline amines, naphthyridine amines, ortetrahydronaphthyridine amines.

Additional examples of small molecule IRMs said to induce interferon(among other things), include purine derivatives (such as thosedescribed in U.S. Pat. Nos. 6,376,501, and 6,028,076), imidazoquinolineamide derivatives (such as those described in U.S. Pat. No. 6,069,149),1H-imidazopyridine derivatives (such as those described in JapanesePatent Application No. 9-255926), benzimidazole derivatives (such asthose described in U.S. Pat. No. 6,387,938), derivatives of a4-aminopyrimidine fused to a five membered nitrogen containingheterocyclic ring (such as adenine derivatives described in U.S. Pat.Nos. 6,376,501; 6,028,076 and 6,329,381; and in InternationalPublication No. WO 02/08595), and certain3-.beta.-D-ribofuranosylthiazolo[4,5-d]pyri-midine derivatives (such asthose described in U.S. Patent Publication No. 2003/0199461).1H-imidazopyridine derivatives (such as those described in U.S. Pat. No.6,518,265 and European Patent Application EP No. 1 256 582)) are said toinhibit TNF and IL-1 cytokines.

Examples of small molecule IRMs that comprise a 4-aminopyrimidine fusedto a five-membered nitrogen-containing heterocyclic ring include adeninederivatives (such as those described in U.S. Pat. Nos. 6,376,501;6,028,076 and 6,329,381; and in International Publication No. WO02/08595).

Examples of particular IRM compounds include2-propyl[1,3]thiazolo[4,5-c]quinolin-4-amine, which is consideredpredominantly a TLR 8 agonist (and not a substantial TLR 7 agonist),4-amino-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol,which is considered predominantly a TLR 7 agonist (and not a substantialTLR 8 agonist), and4-amino-2-(ethoxymethyl)-alpha,alpha.-dimethyl-6,7,8,9-tetrahydro-1H-imidazo[4,5-c]quinolines-1-ethanol,which is a TLR 7 and TLR 8 agonist. In addition to its TLR 7 activity(and TLR 6 activity, but low TLR 8 activity),4-amino-alpha,alpha-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol hasbeneficial characteristics, including that it has a much lower CNSeffect when delivered systemically compared to imiquimod. Other examplesof specific IRM compounds include, e.g.,N-[4-(4-amino-2-butyl-1H-imidazo[4,5-c][1,5]naphthyridin-1-yl)butyl]-N′-cyclohexylurea,2-methyl-1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyridin-4-amine,1-(2-methylpropyl)-1H-imidazo[4,5-c][1,5]naphthyridin-4-amine,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimethylethyl}methanesulfonamide,N-[4-(4-amino-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl]methanesulfonamide,2-methyl-1-[5-(methylsulfonyl)pentyl]-1H-imidazo[4,5-c]quinolin-4-amine,N-[4-(4-amino-2-propyl-1H-imidazo[4,5-c]quinolin-1-yl)butyl]methanesulfonamide,2-butyl-1-[3-(methylsulfonyl)propyl-]-1H-imidazo[4,5-c]quinoline-4-amine,2-butyl-1-{2-[(1-methylethyl)sulfonyl]ethyl}-1H-imidazo-[4,5-c]quinolin-4-amine,N-{2-[4-amino-2-(ethoxymethyl)-1-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimethylethyl}-N′-cyclohexylurea,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[4,5-c]quinolin-1-yl]-1,1-dimethylethyl}cyclohexanecarboxamide,N-{2-[4-amino-2-(ethoxymethyl)-1H-imidazo[-4,5-c]quinolin-1-yl]ethyl}-N′-isopropylurea.Resiquimod,4-amino-2-ethoxymethyl-.alpha.,.alpha.-dimethyl-1H-imidazo[4,5-c]quinoline-1-ethanol,may also be used in certain situations where a combination TLR 7 and TLR8 agonist is desired.

Other IRMs include large biological molecules such as oligonucleotidesequences. Some IRM oligonucleotide sequences contain cytosine-guaninedinucleotides (CpG) and are described, for example, in U.S. Pat. Nos.6,194,388; 6,207,646; 6,239,116; 6,339,068; and 6,406,705. SomeCpG-containing oligonucleotides can include synthetic immunomodulatorystructural motifs such as those described, for example, in U.S. Pat.Nos. 6,426,334 and 6,476,000. Other IRM nucleotide sequences lack CpGand are described, for example, in International Patent Publication No.WO 00/75304. IRMs are delivered agents in embodiments of the transdermaldelivery compositions of the present invention.

Embodiments of the invention are also useful for delivery of compoundsused to facilitate imaging of tissues and organs within the body.Several imaging methods commonly used include Xray, CT scans,ultrasound, and magnetic resonance imaging. Various compounds areadministered to individuals that facilitate the imaging process. Thus,other embodiments are useful for the delivery of diagnostic or contrastcomponents useful in imaging methods now known or later discoveredinclude iohexyl, technetium, Tc99M, sestamibi, iomeprol, gadodiamide,oiversol, iopromide, alsactide, americium, betazole, histamine,mannitol, metyraphone, petagastrin, phentolamine, radioactive B12,gadodiamide, gadopentetic acid, gadoteridol, or perflubron as deliveredagents.

In addition to low molecular weight delivered agents and mediummolecular weight delivered agents, several high molecular weightdelivered agents (e.g., glycoproteins) can be delivered to cells in thebody by using an embodiment of the transdermal delivery composition.Glycoproteins are high molecular weight compounds, which are generallycharacterized as conjugated proteins containing one or moreheterosaccharides as prosthetic groups. The heterosaccharides areusually branched but have a relatively low number of sugar residues,lack a serially repeating unit, and are covalently bound to apolypeptide chain. Several forms of glycoproteins are found in the body.For example, many membrane bound proteins are glycoproteins, thesubstances that fill the intercellular spaces (e.g., extracellularmatrix proteins) are glycoproteins, and the compounds that composecollagens, proteoglycans, mucopolysaccharides, glycosaminoglycans, andground substance are glycoproteins. A delivery system that canadminister glycoproteins to cells of the body has several pharmaceuticaland cosmetic uses, including but not limited to, the restoration of skinelasticity and firmness (e.g., the reduction in the appearance of finelines and wrinkles by transdermal delivery of collagen) and therestoration of flexible and strong joints (e.g., water retention injoints can be increased by transdermal delivery of proteoglycans).

Accordingly, in another aspect of the invention, a transdermal deliverycomposition that can administer a high molecular weight compound (e.g.,a form of collagen or fragment thereof) to cells of the body isprovided. As described above, many different high molecular weightcompounds can be administered by using an embodiment of a transdermaldelivery composition of the invention and the use of a high molecularweight collagen as a delivered agent is intended to demonstrate thatembodiments of the invention can deliver many high molecular weightcompounds to cells of the body.

Collagens exist in many forms and can be isolated from a number ofsources. Additionally, several forms of collagen can be obtainedcommercially (e.g., Brooks Industries Inc., New Jersey). Many lowmolecular weight collagens can be made, for example, by hydrolysis.Several transdermal delivery compositions of the invention can delivercollagens having molecular weights below 6,000 daltons. Additionally,several high molecular weight collagens exist. Some are isolated fromanimal or plant sources and some are synthesized or produced throughtechniques common in molecular biology. Several transdermal deliverycompositions of the invention can deliver collagens having molecularweights of 1,000 daltons to greater than 2,000,000 daltons. That is,embodiments of the transdermal delivery compositions can delivercollagens having molecular weights of less than or equal to or greaterthan 1,000, 1,500, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, 5,000,5,500, 6,000, 7,000, 8,000, 9,000, 10,000, 11,000, 12,000, 13,000,14,000, 15,000, 16,000, 17,000, 18,000, 19,000, 20,000, 21,000, 22,000,23,000, 24,000, 25,000, 26,000, 27,000, 28,000, 29,000, 30,000, 31,000,32,000, 33,000, 34,000, 35,000, 36,000, 37,000, 38,000, 39,000, 40,000,41,000, 42,000, 43,000, 44,000, 45,000, 46,000, 47,000, 48,000, 49,000,50,000, 51,000, 52,000, 53,000, 54,000, 55,000, 56,000, 57,000, 58,000,59,000, 60,000, 61,000, 62,000, 63,000, 64,000, 65,000, 66,000, 67,000,68,000, 69,000, 70,000, 75,000, 80,000, 85,000, 90,000, 95,000, 100,000,125,000, 150,000, 175,000, 200,000, 225,000, 250,000, 275,000, 300,000,350,000, 400,000, 450,000, 500,000, 600,000, 700,000, 800,000, 900,000,1,000,000, 1,500,000, 1,750,000, and 2,000,000 daltons.

In some embodiments, the commercially available collagen “HydrocollEN-55” was provided as the delivered agent and was delivered to cells ofa test subject. This form of collagen is hydrolyzed collagen and has amolecular weight of 2,000 daltons. In another embodiment, thecommercially available “Ichtyocollagene” or marine collagen (Sederma orCroda of Parsippany, N.J.) was provided as the delivered agent and wasdelivered to a test subject. This form of soluble collagen has amolecular weight of greater than 100,000 daltons. In another embodiment,the commercially available collagen “Solu-Coll” was provided as thedelivered agent and was delivered to cells of a test subject. This formof collagen is a soluble collagen having a molecular weight of 300,000daltons. An additional embodiment includes the commercially availablecollagen “Plantsol”, which is obtained from yeast and has a molecularweight of 500,000 daltons. This collagen was also provided as adelivered agent and was delivered to cells of a test subject.

The transdermal delivery compositions that contain a form of collagen orfragment thereof desirably comprise by weight or volume between 0.1% to85.0% of the delivered agent depending on the type and form of thecollagen, its solubility, and the intended application. That is, sometransdermal delivery compositions comprise by weight or volume less thanor equal to or greater than 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%,0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%,1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%,3.75%, 4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%,6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%,9.25%, 9.5%, 9.75%, 10.0%, 10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%,11.75%, 12.0%, 12.25%, 12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%,14.0%, 14.25%, 14.5%, 14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%,18.0%, 18.5%, 19.0%, 19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%,23.0%, 23.5%, 24.0%, 24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%,28.0%, 28.5%, 29.0%, 29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%,33.0%, 33.5%, 34.0%, 34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%,38.0%, 38.5%, 39.0%, 39.5%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%,46.0%, 47.0%, 48.0%, 49.0%, 50.0%, 51%, 52%, 53%, 54%, 55%, 56%, 57%,58%, 59%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%,81%, 82%, 83%, 84%, or 85% collagen or fragment thereof.

For example, embodiments having Hydrocoll-EN55 can comprise by weight orvolume less than or equal to or greater than 1.0%, 1.25%, 1.5%, 1.75%,2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%,4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%,7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, 10.0%,10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%, 12.0%, 12.25%,12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%, 14.25%, 14.5%,14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%,19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%, 23.5%, 24.0%,24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%,29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%, 33.5%, 34.0%,34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%, 38.5%, 39.0%,39.5%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%,49.0%, 50.0%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, or85% Hydrocoll-EN-55.

Embodiments having marine collagen can comprise by weight or volume lessthan or equal to or greater than 1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.25%,2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%, 4.75%, 5.0%,5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%,8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, 10.0%, 10.25%, 10.5%,10.75%, 11.0%, 11.25%, 11.5%, 11.75%, 12.0%, 12.25%, 12.5%, 12.75%,13.0%, 13.25%, 13.5%, 13.75%, 14.0%, 14.25%, 14.5%, 14.75%, 15.0%,15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%, 19.5%, 20.0%,20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%, 23.5%, 24.0%, 24.5%, 25.0%,25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%, 29.5%, 30.0%,30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%, 33.5%, 34.0%, 34.5%, 35.0%,35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%, 38.5%, 39.0%, 39.5%, 40.0%,41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%, 49.0%, 50.0%,51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 69%, 70%, 71%, 72%, 73%,74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, or 85% marinecollagen.

Further, transdermal delivery compositions that contain Solu-Coll cancomprise by weight or volume less than or equal to or greater than 0.1%,0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%,0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%, 1.1%, 1.15%, 1.2%, 1.25%,1.3%, 1.35%, 1.4%, 1.45%, 1.5%, 1.55%, 1.6%, 1.65%, 1.7%, 1.75%, 1.8%,1.85%, 1.9%, 1.95%, or 2.0% Solu-Coll.

Additionally, transdermal delivery compositions that contain Plantsolcan comprise by weight or volume less than or equal to or greater than0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%,0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%, 1.1%, 1.15%, 1.2%,1.25%, 1.3%, 1.35%, 1.4%, 1.45%, 1.5%, 1.55%, 1.6%, 1.65%, 1.7%, 1.75%,1.8%, 1.85%, 1.9%, 1.95%, 2.0%, 2.1%, 2.15%, 2.2%, 2.25%, 2.3%, 2.35%,2.4%, 2.45%, 2.5%, 2.55%, 2.6%, 2.65%, 2.7%, 2.75%, 2.8%, 2.85%, 2.9%,2.95%, 3.0%, 3.1%, 3.15%, 3.2%, 3.25%, 3.3%, 3.35%, 3.4%, 3.45%, 3.5%,3.55%, 3.6%, 3.65%, 3.7%, 3.75%, 3.8%, 3.85%, 3.9%, 3.95%, or 4.0%Plantsol.

In other embodiments of the invention, a transdermal deliverycomposition that can provide a collagen solution comprising two or moreforms of collagen (e.g., Hydro-Coll EN-55, marine collagen, Solu-coll,or Plantsol) is provided. The transdermal delivery compositions thatinclude two or more forms of collagen desirably comprise an amount ofdelivered agent that can be included in a delivered agent having thespecific type of collagen by itself. For example, if the mixture ofdelivered agents comprises Hydro-Coll EN55, the amount of Hydro-CollEN55 in the transdermal delivery composition can comprise by weight orvolume less than or equal to or greater than 1.0%, 1.25%, 1.5%, 1.75%,2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%,4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%,7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, 10.0%,10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%, 12.0%, 12.25%,12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%, 14.25%, 14.5%,14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%,19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%, 23.5%, 24.0%,24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%,29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%, 33.5%, 34.0%,34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%, 38.5%, 39.0%,39.5%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%,49.0%, 50.0%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, or85% Hydrocoll-EN-55.

If the mixture of delivered agents has marine collagen, then the amountof marine collagen in the delivery system can comprise by weight orvolume less than or equal to or greater than 1.0%, 1.25%, 1.5%, 1.75%,2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%,4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%,7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, 10.0%,10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%, 12.0%, 12.25%,12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%, 14.25%, 14.5%,14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%,19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%, 23.5%, 24.0%,24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%,29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%, 33.5%, 34.0%,34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%, 38.5%, 39.0%,39.5%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%, 46.0%, 47.0%, 48.0%,49.0%, 50.0%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 69%, 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, or85% marine collagen. Similarly if the mixture of delivered agents hasSolu-coll, then the amount of Solu-coll in the delivery system cancomprise by weight or volume less than or equal to or greater than 0.1%,0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%,0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%, 1.1%, 1.15%, 1.2%, 1.25%,1.3%, 1.35%, 1.4%, 1.45%, 1.5%, 1.55%, 1.6%, 1.65%, 1.7%, 1.75%, 1.8%,1.85%, 1.9%, 1.95%, or 2.0% or Solu-Coll. Further, if the mixture ofdelivered agents has Plantsol, then the amount of Plantsol in thedelivery system can comprise by weight or volume less than or equal toor greater than 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%,0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%,1.1%, 1.15%, 1.2%, 1.25%, 1.3%, 1.35%, 1.4%, 1.45%, 1.5%, 1.55%, 1.6%,1.65%, 1.7%, 1.75%, 1.8%, 1.85%, 1.9%, 1.95%, 2.0%, 2.1%, 2.15%, 2.2%,2.25%, 2.3%, 2.35%, 2.4%, 2.45%, 2.5%, 2.55%, 2.6%, 2.65%, 2.7%, 2.75%,2.8%, 2.85%, 2.9%, 2.95%, 3.0%, 3.1%, 3.15%, 3.2%, 3.25%, 3.3%, 3.35%,3.4%, 3.45%, 3.5%, 3.55%, 3.6%, 3.65%, 3.7%, 3.75%, 3.8%, 3.85%, 3.9%,3.95%, or 4.0% Plantsol.

Additionally, modified or stabilized collagens or collagen derivativesare contemplated for use in some of the embodiments described herein.Particularly preferred are collagens that are resistant to proteases.Recombinant engineering can be used to generate collagens or fragmentsthereof that lack protease cleavage sites for example. Resistantcollagens or fragments thereof can also be prepared by incorporatingD-amino acids in synthetically prepared collagens or fragments thereof.Cross-linked collagens can also be used. (See e.g., Charulatha,Biomaterials February;24(5):759-67 (2003)). Still further, amidatedcollagen or collagen fragments can be prepared using synthetic chemistryand these collagen derivatives can be mixed with an ethoxylated oil withor without water or alcohol so as to form a transdermal deliverycomposition containing collagen. Several techniques to create synthetic,recombinant, or cross-linked collagens are known to those of skill inthe art and many are commercially available.

Still further, protease resistant fragments of collagen can be preparedand isolated using conventional techniques. By one approach, marinecollagen, procollagen, or collagen obtained from human placenta isincubated with bovine serum, pepsin, or bacterial collagenase for onehour and the preparation is then separated by gel electrophoresis, sizeexclusion, reverse phase, or ionic exchange chromatography (e.g., FPLCor HPLC). Protease resistant fragments of collagen (e.g., 15 kDa or 30kDa; see e.g., Tasab et al., JBC 277(38):35007 (2002) or 38 kDa seee.g., Odermatt et al., Biochem J May 1; 211(2):295-302 (1983) both ofwhich are herein expressly incorporated by reference in theirentireties) are separated from the hydrolytic products and thesefragments are isolated from the column and concentrated (e.g., centriconfilters) or lyophilized using conventional techniques. The proteaseresistant fragments of collagen are then incorporated into a transdermaldelivery composition, as described herein. Alternatively, the proteaseresistant domain of collagen can be prepared synthetically or obtainedcommercially (e.g., pepsinized collagens can also be obtained fromChemicon of Temecula, Calif.).

An additional delivered agent that can be included in a transdermaldelivery composition is Etioline (Sederma or Croda of Parsippany, N.J.).Etioline is a tyrosinase inhibitor made from the extract Mitracarpe andbearberry that effectively whitens the skin. Formulations of atransdermal delivery composition described herein containing Etioline(e.g., at 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%,15%, 16%, 17%, 18%, 19%, or 20%) are also embodiments of the invention.Another skin brightening or whitening formulation of a transdermaldelivery composition comprises Melaslow (Sederma of Parsippany, N.J.).Melaslow is an extract made from Citrus reticulate Blanco var. Unshiu.Melaslow is also an inhibitor of melanogenesis and formulations of atransdermal delivery composition described herein containing Melaslow(e.g., at 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%,15%, 16%, 17%, 18%, 19%, or 20%) are also embodiments of the invention.An additional delivered agent that can be included in a transdermaldelivery composition is Matrixyl (Sederma or Croda of Parsippany, N.J.).Matrixyl is a compound comprising the peptide KTTKS (SEQ. ID. NO:2),which has been shown to stimulate collagen synthesis. See Katayama etal., J. Biol. Chem. 268, 9941 (1993). Formulations of a transdermaldelivery composition described herein containing Matrixyl or the peptideKTTKS (SEQ. ID. NO:2) (e.g., at 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20%) are alsoembodiments of the invention. The section below describes themanufacture and use of several penetration enhancers that deliver bothlow and high molecular weight molecules to cells of the body.

Penetration Enhancers

A penetration enhancer included in many embodiments of the invention iscomprised of two components—a hydrophobic component and a hydrophiliccomponent. Desirably, the hydrophobic component comprises a polyethercompound, such as an ethoxylated fatty moiety, preferably, anethoxylated oil, such as vegetable, nut, synthetic, or animal oil, whichhas the ability to reduce the surface tension of materials that aredissolved into it. Not wanting to be tied to any particular mechanism ormode of action and offered only to expand the knowledge in the field, itis contemplated that the attachment of poly (ethylene oxide) to thecomponents of a particular oil occurs not on a particular functionalgroup but rather the polyethylene oxide chains begin to grow fromunsaturated C═C bonds and from the occasional glycerol unit. Because anethoxylated oil, such as ethoxylated macadamia nut oil, is a mixture ofvarious fatty acids, fatty alcohols, and fatty amines, the components ofthe oil may have varying amounts of ethoxylation. Accordingly,measurements of ethoxylation/molecule (e.g., 16 ethoxylations/molecule)are an average of the amount of ethoxylation present on the componentsof the oil rather than on any specific component itself.

Preferred ethoxylated oils can be obtained or created from, for example,macadamia nut oil, meadowfoam, castor oil, jojoba oil, corn oil,sunflower oil, sesame oil, and emu oil. Many of these oils arecommercially available from Floratech of Gilbert, Ariz. or othersuppliers. Alternatively, ethoxylated oils can be prepared by reactingthe oil with ethylene oxide. Pure carrier oils that are suitable forethoxylation so as to create a penetration enhancer for use with thetransdermal delivery compositions described herein are included inTABLES 3-17 and can be obtained from Esoteric oils Pty. Ltd., PretoriaSouth Africa. TABLES 3-17 also list the component fatty acids of theseoils, all of which are individually suitable for ethoxylation andincorporation into an embodiment of a transdermal delivery composition.That is, it is contemplated that ethoxylated fatty acids, ethoxylatedfatty alcohols, and ethoxylated fatty amines, in particular ethoxylatedfatty acids, ethoxylated fatty alcohols, and ethoxylated fatty aminesthat contain 12, 13, 14, 15, 16, 17, 18, or 19 ethoxylations aresuitable penetration enhancers for use in the transdermal deliverycompositions described herein. These ethoxylated oil components can beused individually as penetration enhancers or as supplements to otherpenetration enhancers (e.g., ethoxylated macadamia nut oil).

TABLE 3 MACADAMIA NUT OIL Fatty acids Range Myristic C14 0.6-1.6%Palmitic C16  7.0-11.0% Palmitoleic C16:1 18.0-25.0% Stearic C182.0-4.0% Oleic C18:1 55.0-62.0% Linoleic C18:2 1.0-4.0% Arachidic C202.0-4.0% Eicosenoic C20:1 2.0-4.0%

TABLE 4 APRICOT KERNEL OIL Fatty acids Range Typical Palmitic C16:03.0-6.0% 4.28% Palmitoleic C16:1 trace-1.4% 0.70% Stearic C18:0trace-2.0% 1.12% Oleic C18:1 55.0-70.0% 69.62% Linoleic C18:2 20.0-35.0%23.34% Linolenic C18:3 trace-1.0% 0.22% Eicosenoic C20:1 trace-1.0%0.18%

TABLE 5 AVOCADO OIL Fatty acids Range Typical Palmitic C16:0 12.0-20.0%14.25% Palmitoleic C16:1  2.0-10.0%  5.84% Stearic C18:0 0.1-2.0%  0.1%Oleic C18:1 55.0-75.0%  65.4% Linoleic C18:2  9.0-17.0% 14.74% LinolenicC18:3 0.1-2.0%  0.8%

TABLE 6 EVENING PRIMROSE OIL Fatty acids Range Typical Palmitic C16:05.5-7.0% 5.9% Stearic C18:0 1.5-2.5% 1.7% Oleic C18:1  5.0-11.0% 5.8%Linoleic C18:2 70.0-77.0% 75.1%  Gamma C18:3  9.0-10.9% 10.6%  LinolenicAlpha C18:3 1.0% max 0.4% Linolenic Icosanoic C20:0 1.0% max 0.2%Icosenoic C20:1 1.0% max .01%

TABLE 7 GRAPE SEED OIL Fatty acids Range Typical Palmitic C16:0 6.0-9.0%6.5% Palmitoleic C16:1 less 1% 0.2% Stearic C18:0 3.0-6.0% 3.7% OleicC18:1 12.0-25.0% 23.4% Linoleic C18:2 60.0-75.0% 65.3% Alpha C18:3 lessthan 1.5% 0.2% Linolenic Icosanoic C20:0 less than 0.5% 0.2% IcosenoicC20:1 less than 0.5% 0.2% Docosanoic C22:0 less than 0.3% 0.2%

TABLE 8 HAZELNUT OIL Fatty acids Range Palmitic C16:0 4.0-8.0%Palmitoleic C16:1 0.1-0.6% Stearic C18:0 1.5-3.5% Oleic C18:1 68.0-85.0%Linoleic C18:2  7.0-15.0% Linolenic C18:3 0.1-0.5% Arachidic C20:00.1-0.5% Gadoleic C20:1 0.1-0.3% Behenic C22:0 3.0% MAX

TABLE 9 JOJOBA OIL Fatty acids Range Palmitic C16:0 3.0% max PalmitoleicC16:1 1.0% max Stearic C18:0 1.0% max Oleic C18:1  5.0-15.0% LinoleicC18:2 5.0% max Linolenic C18:3 1.0% max Arachidic C20:0 0.5% maxEicosenoic C20:1 65.0-80.0% max Behenic C22:0 0.5% max Erucic C22:110.0-20.0% max Lignoceric C24:0 5.0% max

TABLE 10 OLIVE OIL Fatty acids Range Palmitic C16:0 5.0-12.0%Palmitoleic C16:1 1.0% max Stearic C18:0 3.5% max Oleic C18:165.0-80.0%  Linoleic C18:2 6.0-25.0% Linolenic C18:3 1.0% max ArachidicC20:0 0.6% max Gadoleic C20:1 0.5% max Behenic C22:0 0.3% max ErucicC22:1 0.2% max

TABLE 11 PUMPKIN SEED OIL Fatty acids Range Palmitic C16:0  6.0-21.0%Stearic C18:0 3.0-8.0% Oleic C18:1 24.0-41.0% Linoleic C18:2 42.0-60.0%Linolenic C18:3 2.0% max Others 2.0% max

TABLE 12 ROSE HIP OIL Fatty acids Range Mirystic C14:0 0.0-0.3% PalmiticC16:0 3.4-4.4% Palmitoleic C16:1  0.1-0.18% Stearic C18:0 1.5-2.5% OleicC18:1 14.0-16.0% Linoleic C18:2 43.0-46.0% Linolenic C18:3 31.0-34.0%Arachidic C20:0 0.1-0.9% Gadoleic C20:1 0.0-0.5% Eicosenoic C20:20.0-0.5% Behenic C22:0 0.1-0.4%

TABLE 13 SAFFLOWER OIL Fatty acids Range Palmitic C16:0 4.0-9.0%Palmitoleic C16:1 Trace Stearic C18:0 trace-2.5% Oleic C18:1 72.0-80.0%Linoleic C18:2 12.0-16.0% Linolenic C18:3 trace-0.5%

TABLE 14 SESAME OIL Fatty acids Range Palmitic C16:0  7.0-12.0%Palmitoleic C16:1 trace-0.5% Stearic C18:0 3.5-6.0% Oleic C18:135.0-50.0% Linoleic C18:2 35.0-50.0% Linolenic C18:3 trace-1.0%Eicosenoic C20:1 trace-1.0%

TABLE 15 SUNFLOWER OIL Fatty acids Range Palmitic C16:0 5.8% PalmitoleicC16:1 0.1% Stearic C18:0 3.9% Oleic C18:1 15.9% Linoleic C18:2 71.7%Alpha Linolenic C18:3 0.6% Gamma Linolenic C18:3 0.1% Arachidic C20:00.3% Gadoleic C20:1 0.2% Tetracosanoic C24:0 0.5% Behenic C22:0 0.7%

TABLE 16 WALNUT OIL Fatty acids Range Typical Palmitic C16:0 5.0-8.0%6.0% Palmitoleic C16:1 less than 1.0% 0.1% Stearic C18:0 3.0-7.0% 4.0%Oleic C18:1 25.0-35.0% 29.8% Linoleic C18:2 45.0-60.0% 58.5% Alpha C18:3less than 0.8% 0.4% Linolenic Arachidic C20:0 less than 0.5% 0.3%Eicosenoic C20:1 less than 0.5% 0.2%

TABLE 17 WHEAT GERM OIL Fatty acids Range Typical Palmitic C16:011.0-16.0% 12.5% Palmitoleic C16:1 1.0% max 0.2% Stearic C18:0 2.0-6.0%2.5% Oleic C18:1 12.0-39.0% 27.3% Linoleic C18:2 30.0-57.0% 53.7%Linolenic C18:3  2.0-10.0% 3.0% Arachidic C20:0 1.0% max 0.4% GadoleicC20:1 0.5% max 0.2% Behenic C22:0 1.0% max 0.1%

In some embodiments, an ethoxylated oil comprises a molar ratio ofethylene oxide:oil of 35:1. A 99% pure ethylene oxide/castor oil havingsuch characteristics can be obtained commercially (BASF) or such anethoxylated compound can be synthesized using conventional techniques.In other embodiments, the ethoxylated oil is itself the penetrationenhancer. That is, it has been discovered that oils that have beenethoxylated 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19ethoxylations/molecule are sufficiently hydrophobic and sufficientlyhydrophilic to allow for transdermal delivery of a variety of deliveredagents without water, alcohol, or an aqueous adjuvant. Although theethoxylated oil can comprise at least 20-25 ethoxylations per moleculeor more, preferably, the ethoxylated lipid comprises less than 20ethoxylations per molecule, e.g., 19, 18, 17, 16, 15, 14, 13, 12, 11, or10 ethoxylations per molecule.

By using a light, ethoxylated oil (e.g., macadamia nut oil containingapproximately 16 ethoxylations/molecule) efficient transdermal deliveryof high molecular weight collagen was observed in the absence of AloeVera and alcohol. Formulations of a transdermal delivery compositionthat contain Aloe Vera and an oil with 20-30 ethoxylations/molecule arenot as effective as formulations of a transdermal delivery compositionthat contain an oil with 10-19 ethoxylations/molecule (e.g., 16ethoxylations/molecule) but lacking Aloe Vera and alcohol. A greaterreduction of fine lines and wrinkles was observed with a transdermaldelivery composition composed of macadamia nut oil (16ethoxylations/molecule) and water as compared with a transdermaldelivery composition composed of castor oil (25 ethoxylations/molecule),water, alcohol, and Aloe Vera, for example.

Unexpectedly, it was discovered that a reduction in the number ofethoxylations on a light oil produced a superior transdermal deliveryproduct. This was unexpected because as the amount of ethoxylations on amolecule of oil decreases its miscibility with the aqueous components ofthe delivery system decreases. Surprisingly, formulations containing10-19 ethoxylations/molecule were not only miscible but provided veryefficient transdermal delivery in the absence of Aloe Vera.

Desirable compounds often found in ethoxylated oils that are beneficialfor some embodiments and methods described herein areglycerol-polyethylene glycol ricinoleate, the fatty esters ofpolyethylene glycol, polyethylene glycol, and ethoxylated glycerol. Someof these desirable compounds exhibit hydrophilic properties and thehydrophilic-lipophilic balance (HLB) is preferably maintained between 10and 18. Any number of methods have been devised to characterize HLB, butperhaps the most widely used is the octanol/water coefficient. (SeeCalculating log Poct from Structures”, by Albert J. Leo, ChemicalReviews, vol 93, pp 1281).

Accordingly, some of the components of the oils in the table above andrelated fatty acids, fatty alcohols, and fatty amines can be ethoxylatedand used as a penetration enhancer or to enhance another penetrationenhancer (e.g., ethoxylated macadamia nut oil). For example, someembodiments comprise a penetration enhancer that consists of, consistsessentially of, or comprises ethoxylated palmitoleic acid, ethoxylatedoleic acid, ethoxylated gondoic acid, or ethoxylated erucic acid. Thesecompounds can be prepared synthetically or isolated or purified fromoils that contain large quantities of these fatty acids and thesynthesized, isolated, or purified fatty acids can then be reacted withethylene oxide.

That is, a transdermal delivery composition of the invention cancomprise a penetration enhancer that contains, for example, ethoxylatedpalmitoleic acid, ethoxylated oleic acid, ethoxylated gondoic acid, orethoxylated erucic acid, wherein the amount of one or more of the fattyacids is at least, less than, more than, or an amount equal to 0.1%,0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.55%, 0.6%, 0.65%,0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%, 1.25%, 1.5%, 1.75%, 2.0%,2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%, 4.0%, 4.25%, 4.5%, 4.75%,5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%, 6.75%, 7.0%, 7.25%, 7.5%,7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%, 9.5%, 9.75%, 10.0%, 10.25%,10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%, 12.0%, 12.25%, 12.5%,12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%, 14.25%, 14.5%, 14.75%,15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%, 18.5%, 19.0%, 19.5%,20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%, 23.5%, 24.0%, 24.5%,25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%, 28.5%, 29.0%, 29.5%,30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%, 33.5%, 34.0%, 34.5%,35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%, 38.5%, 39.0%, 39.5%,40.0%, 40.25%, 40.5%, 40.75%, 41.0%, 41.25%, 41.5%, 41.75%, 42.0%,42.25%, 42.5%, 42.75%, 43.0%, 43.25%, 43.5%, 43.75%, 44.0%, 44.25%,44.5%, 44.75%, 45.0%, 45.25%, 45.5%, 45.75%, 46.0%, 46.25%, 46.5%,46.75%, 47.0% 47.25%, 47.5%, 47.75%, 48.0%, 48.25%, 48.5%, 48.75%,49.0%, 49.25%, 49.5%, 49.75%, 50.0%, 50.25%, 50.5%, 50.75%, 51.0%,51.25%, 51.5%, 51.75%, 52.0%, 52.25%, 52.5%, 52.75%, 53.0%, 53.25%,53.5%, 53.75%, 54.0%, 54.5%, 54.0%, 54.5%, 55.0%, 55.5%, 56.0%, 56.5%,57.0%, 57.5%, 58.0%, 58.5%, 59.0%, 59.5%, 60.0%, 60.5%, 61.0%, 61.5%,62.0%, 62.5%, 63.0%, 63.5%, 64.0%, 64.5%, 65.0%, 65.5%, 66.0%, 66.5%,67.0%, 67.5%, 68.0%, 68.5%, 69.0%, 69.5%, 70.0%, 70.5%, 71.0%, 71.5%,72.0%, 72.5%, 73.0%, 73.5%, 74.0%, 74.5%, 75.0%, 75.5%, 76.0%, 76.5%,77.0%, 77.5%, 78.0%, 78.5%, 79.0%, 79.5%, 80.0%, 80.5%, 81%, 81.5%, 82%,82.5%, 83%, 83.5%, 84%, 84.5%, 85% 85.5%, 86%, 86.5%, 87%, 87.5%, 88%,88.5%, 89%, 89.5%, 90%, 90.5%, 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, 94%,94.5%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, or 100% ofthe total fatty acid content in the composition. In some embodiments,more than one ethoxylated compound is added or another hydrophobiccompound is added (e.g., Y-Ling-Y-Lang oil; Young Living Essential Oils,Lehl, Utah)) to balance or enhance the penetration enhancer. Preferredembodiments include ethoxylated macadamia nut oil that has beensupplemented with ethoxylated palmitoleic acid, ethoxylated oleic acid,ethoxylated gondoic acid, or ethoxylated erucic acid.

Depending on the type of delivered agent and the intended application,the amount of ethoxylated lipid(s) in the delivery system can vary. Forexample, delivery systems of the invention can comprise between 0.1% and99% by weight or volume ethoxylated compound(s). That is, embodiments ofthe invention can comprise by weight or volume at least, less than, orequal to or greater than 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%,0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%,1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%,3.75%, 4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%,6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%,9.25%, 9.5%, 9.75%, 10.0%, 10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%,11.75%, 12.0%, 12.25%, 12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%,14.0%, 14.25%, 14.5%, 14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%,18.0%, 18.5%, 19.0%, 19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%,23.0%, 23.5%, 24.0%, 24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%,28.0%, 28.5%, 29.0%, 29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%,33.0%, 33.5%, 34.0%, 34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%,38.0%, 38.5%, 39.0%, 39.5%, 40.0%, 41%, 42%, 43%, 44%, 45%, 46%, 47%,48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%,62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%,76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% ethoxylatedlipid(s), preferably an ethoxylated oil or fatty acid or combination offatty acids.

The hydrophilic component of the penetration enhancer can comprise analcohol, a non-ionic solubilizer, or an emulsifier. Compounds such asethylene glycol, propylene glycol, dimethyl sulfoxide (DMSO), dimethylpolysiloxane (DMPX), oleic acid, caprylic acid, isopropyl alcohol,1-octanol, ethanol (denatured or anhydrous), and other pharmaceuticalgrade or absolute alcohols with the exception of methanol can be used.Preferred embodiments comprise an alcohol (e.g., absolute isopropylalcohol), which is commercially available. As above, the amount ofhydrophilic component in the penetration enhancer depends on the type ofthe delivered agent and the intended application. The hydrophiliccomponent of a penetration enhancer of the invention can comprisebetween 0.1% and 50% by weight or volume. That is, a delivery system ofthe invention can comprise by weight or volume at least, less than orequal to or greater than 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%,0.45%, 0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%,1.0%, 1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%,3.75%, 4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%,6.5%, 6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%,9.25%, 9.5%, 9.75%, 10.0%, 10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%,11.75%, 12.0%, 12.25%, 12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%,14.0%, 14.25%, 14.5%, 14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%,18.0%, 18.5%, 19.0%, 19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%,23.0%, 23.5%, 24.0%, 24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%,28.0%, 28.5%, 29.0%, 29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%,33.0%, 33.5%, 34.0%, 34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%,38.0%, 38.5%, 39.0%, 39.5%, 40.0%, 41.0%, 42.0%, 43.0%, 44.0%, 45.0%,46.0%, 47.0%, 48.0%, 49.0%, or 50.0% hydrophilic component.

In addition to a delivered agent and penetration enhancer, thetransdermal delivery compositions described herein can comprise anaqueous adjuvant. The section below describes the incorporation ofaqueous adjuvants in formulations of transdermal delivery compositions,in particular, Aloe Vera, which can enhance the delivery of both low andhigh molecular weight molecules to the skin cells of the body.

Aqueous Adjuvants

Several embodiments of the transdermal delivery composition describedherein comprise an aqueous adjuvant such as Aloe Vera juice or water orboth. The term “Aloe” refers to the genus of South African plants of theLiliaceae family, of which the Aloe barbadensis plant is a species. Aloeis an intricate plant, which contains many biologically activesubstances. (Cohen, et al. in Wound Healing/Biochemical and ClinicalAspects, 1st ed. WB Saunders, Philadelphia (1992)). Over 300 species ofAloe are known, most of which are indigenous to Africa. Studies haveshown that the biologically active substances are located in threeseparate sections of the Aloe leaf—a clear gel fillet located in thecenter of the leaf, in the leaf rind or cortex of the leaf and in ayellow fluid contained in the pericyclic cells of the vascular bundles,located between the leaf rind and the internal gel fillet, referred toas the latex. Historically, Aloe products have been used indermatological applications for the treatment of burns, sores and otherwounds. These uses have stimulated a great deal of research inidentifying compounds from Aloe plants that have clinical activity,especially anti-inflammatory activity. (See e.g., Grindlay and Reynolds(1986) J. of Ethnopharmacology 16:117-151; Hart, et al. (1988) J. ofEthnopharmacology 23:61-71). As a result of these studies there havebeen numerous reports of Aloe compounds having diverse biologicalactivities, including anti-tumor activity, anti-gastric ulcer,anti-diabetic, anti-tyrosinase activity, (See e.g., Yagi, et al. (1977)Z. Naturforsch 32c:731-734), and antioxidant activity (InternationalApplication Serial No. PCT/US95/07404).

Recent research has also shown that Aloe Vera, a term used to describethe extract obtained from processing the entire leaf, isolated from theAloe Vera species of Aloe, can be used as a vehicle for deliveringhydrocortisone, estradiol, and testosterone propionate. (See Davis, etal, JAPMA 81:1 (1991) and U.S. Pat. No. 5,708,038 to Davis)). As setforth in Davis (U.S. Pat. No. 5,708,308), one embodiment of “Aloe Vera”can be prepared by “whole-leaf processing” of the whole leaf of the Aloebarbadensis plant. Briefly, whole leaves obtained from the Aloebarbadensis plant are ground, filtered, treated with cellulase(optional) and activated carbon and lyophilized. The lyophilized powderis then reconstituted with water prior to use.

Aloe Vera can be obtained commercially through Aloe Laboratories, forexample. In other embodiments, the Aloe Vera is made as follows. First,the leaves are manually harvested. Next, the leaves are washed withwater and the thorns on both ends are cut. The leaves are thenhand-filleted so as to extract the inner part of the leaf. The inner gelis passed through a grinder and separator to remove fiber from the gel.Then the gel is put into a pasteurizing tank where L-Ascorbic Acid(Vitamin C) and preservatives are added. The gel is pasteurized at 85°C. for 30 minutes. After pasteurization, the gel is put into a holdingtank for about one or two days, after which the gel is sent through a 2micron filter. Finally, the gel is cooled down through a heat exchangerand stored in a steamed, sanitized and clean 55 gallon drum. The abovedescribed sources and manufacturing methods of Aloe Vera are given asexamples and not intended to limit the scope of the invention. One ofordinary skill in the art will recognize that Aloe Vera is a well knownterm of art, and that Aloe Vera is available from various sources andmanufactured according to various methods.

Absolute Aloe Vera (100% pure) can also be obtained from commercialsuppliers (Lily of the Desert, Irving, Tex.). Aloe Vera juice, preparedfrom gel fillet, has an approximate molecular weight of 200,000 to1,400,000 daltons. Whole leaf Aloe Vera gel has a molecular weight of200,000 to 3,000,000 depending on the purity of the preparation.Although, preferably, the embodiments of the invention having Aloe Veracomprise Aloe Vera juice, other extracts from a member of the Liliaceaefamily can be used (e.g., an extract from another Aloe species).

Transdermal delivery compositions having Aloe Vera can comprise between0.1% to 85.0% by weight or volume Aloe Vera. That is, embodiments of theinvention can comprise by weight or volume at least, less than or equalto or greater than 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%,0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%,1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%,4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%,6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%,9.5%, 9.75%, 10.0%, 10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%,12.0%, 12.25%, 12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%,14.25%, 14.5%, 14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%,18.5%, 19.0%, 19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%,23.5%, 24.0%, 24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%,28.5%, 29.0%, 29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%,33.5%, 34.0%, 34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%,38.5%, 39.0%, 39.5%, 40.0%, 40.25%, 40.5%, 40.75%, 41.0%, 41.25%, 41.5%,41.75%, 42.0%, 42.25%, 42.5%, 42.75%, 43.0%, 43.25%, 43.5%, 43.75%,44.0%, 44.25%, 44.5%, 44.75%, 45.0%, 45.25%, 45.5%, 45.75%, 46.0%,46.25%, 46.5%, 46.75%, 47.0% 47.25%, 47.5%, 47.75%, 48.0%, 48.25%,48.5%, 48.75%, 49.0%, 49.25%, 49.5%, 49.75%, 50.0%, 50.25%, 50.5%,50.75%, 51.0%, 51.25%, 51.5%, 51.75%, 52.0%, 52.25%, 52.5%, 52.75%,53.0%, 53.25%, 53.5%, 53.75%, 54.0%, 54.5%, 54.0%, 54.5%, 55.0%, 55.5%,56.0%, 56.5%, 57.0%, 57.5%, 58.0%, 58.5%, 59.0%, 59.5%, 60.0%, 60.5%,61.0%, 61.5%, 62.0%, 62.5%, 63.0%, 63.5%, 64.0%, 64.5%, 65.0%, 65.5%,66.0%, 66.5%, 67.0%, 67.5%, 68.0%, 68.5%, 69.0%, 69.5%, 70.0%, 70.5%,71.0%, 71.5%, 72.0%, 72.5%, 73.0%, 73.5%, 74.0%, 74.5%, 75.0%, 75.5%,76.0%, 76.5%, 77.0%, 77.5%, 78.0%, 78.5%, 79.0%, 79.5%, 80.0%, 80.5%,81%, 81.5%, 82%, 82.5%, 83%, 83.5%, 84%, 84.5%, and 85% Aloe Vera.

The amount of water in the delivery system generally depends on theamount of other reagents (e.g., delivered agent, penetration enhancer,and other aqueous adjuvants or fillers). Although water is used as thesole aqueous adjuvant in some embodiments, preferred embodiments useenough water to make the total volume of a particular preparation of adelivery system such that the desired concentrations of reagents in thepenetration enhancer, aqueous adjuvant, and delivered agent areachieved. Suitable forms of water are deionized, distilled, filtered orotherwise purified. Clearly, however, any form of water can be used asan aqueous adjuvant.

Transdermal delivery compositions having water can comprise between 0.1%to 85.0% by weight or volume water. That is, embodiments of theinvention can comprise by weight or volume at least, less than or equalto or greater than 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%,0.5%, 0.55%, 0.6%, 0.65%, 0.7%, 0.75%, 0.8%, 0.85%, 0.9%, 0.95%, 1.0%,1.25%, 1.5%, 1.75%, 2.0%, 2.25%, 2.5%, 2.75%, 3.0%, 3.25%, 3.5%, 3.75%,4.0%, 4.25%, 4.5%, 4.75%, 5.0%, 5.25%, 5.5%, 5.75%, 6.0%, 6.25%, 6.5%,6.75%, 7.0%, 7.25%, 7.5%, 7.75%, 8.0% 8.25%, 8.5%, 8.75%, 9.0%, 9.25%,9.5%, 9.75%, 10.0%, 10.25%, 10.5%, 10.75%, 11.0%, 11.25%, 11.5%, 11.75%,12.0%, 12.25%, 12.5%, 12.75%, 13.0%, 13.25%, 13.5%, 13.75%, 14.0%,14.25%, 14.5%, 14.75%, 15.0%, 15.5%, 16.0%, 16.5%, 17.0%, 17.5%, 18.0%,18.5%, 19.0%, 19.5%, 20.0%, 20.5%, 21.0%, 21.5%, 22.0%, 22.5%, 23.0%,23.5%, 24.0%, 24.5%, 25.0%, 25.5%, 26.0%, 26.5%, 27.0%, 27.5%, 28.0%,28.5%, 29.0%, 29.5%, 30.0%, 30.5%, 31.0%, 31.5%, 32.0%, 32.5%, 33.0%,33.5%, 34.0%, 34.5%, 35.0%, 35.5%, 36.0%, 36.5%, 37.0%, 37.5%, 38.0%,38.5%, 39.0%, 39.5%, 40.0%, 40.25%, 40.5%, 40.75%, 41.0%, 41.25%, 41.5%,41.75%, 42.0%, 42.25%, 42.5%, 42.75%, 43.0%, 43.25%, 43.5%, 43.75%,44.0%, 44.25%, 44.5%, 44.75%, 45.0%, 45.25%, 45.5%, 45.75%, 46.0%,46.25%, 46.5%, 46.75%, 47.0% 47.25%, 47.5%, 47.75%, 48.0%, 48.25%,48.5%, 48.75%, 49.0%, 49.25%, 49.5%, 49.75%, 50.0%, 50.25%, 50.5%,50.75%, 51.0%, 51.25%, 51.5%, 51.75%, 52.0%, 52.25%, 52.5%, 52.75%,53.0%, 53.25%, 53.5%, 53.75%, 54.0%, 54.5%, 54.0%, 54.5%, 55.0%, 55.5%,56.0%, 56.5%, 57.0%, 57.5%, 58.0%, 58.5%, 59.0%, 59.5%, 60.0%, 60.5%,61.0%, 61.5%, 62.0%, 62.5%, 63.0%, 63.5%, 64.0%, 64.5%, 65.0%, 65.5%,66.0%, 66.5%, 67.0%, 67.5%, 68.0%, 68.5%, 69.0%, 69.5%, 70.0%, 70.5%,71.0%, 71.5%, 72.0%, 72.5%, 73.0%, 73.5%, 74.0%, 74.5%, 75.0%, 75.5%,76.0%, 76.5%, 77.0%, 77.5%, 78.0%, 78.5%, 79.0%, 79.5%, 80.0%, 80.5%,81%, 81.5%, 82%, 82.5%, 83%, 83.5%, 84%, 84.5%, and 85% water. Inaddition to the aforementioned compositions, methods of making and usingthe transdermal delivery compositions are described in the followingsection.

Preparing Transdermal Delivery Compositions

In general, transdermal delivery compositions are prepared by combiningan ethoxylated fatty moiety or a penetration enhancer with a deliveredagent and, optionally, an aqueous adjuvant. Depending on the solubilityof the delivered agent, the delivered agent can be solubilized in eitherthe hydrophobic or hydrophilic components of the penetration enhancer.In some formulations, (e.g., formulations containing oil solubledelivered agents such as steroid hormones), the delivered agent readilydissolves in the ethoxylated oil without water, alcohol, or an aqueousadjuvant. In other formulations, the delivered agent (e.g., an NSAID orcollagen or fragments thereof) readily dissolves in water, which is thenmixed with the ethoxylated oil. Additionally, some delivered agents canbe solubilized in the aqueous adjuvant prior to mixing with thepenetration enhancer. Desirably, the pH of the mixture is maintainedbetween 3 and 11 and preferably between 5 and 9. That is, duringpreparation and after preparation the pH of the solution is desirablymaintained at less than, more than, at least, or equal to 3.0, 3.25,3.5, 3.75, 4.0, 4.25, 4.5, 4.75, 5.0, 5.25, 5.5, 5.75, 6, 6.25, 6.5,6.75, 7.0, 7.25, 7.5, 7.75, 8.0, 8.25, 8.5, 8.75, 9.0, 9.25, 9.5, 9.75,10.0, 10.25, 10.5, 10.75, or 11.0.

Several physical mixing techniques can be employed to help the deliverysystem coalesce. For example, a magnetic stir plate and bar can be used,however, the speed of stirring is preferably minimized so as not todrive air into the mixture and/or destroy the delivered agent (e.g.,when the delivered agent is a peptide or a protein). Additionally, arocker can be used to bring components of the delivery system together.Heat can also be applied to help coalesce the mixture but desirably, thetemperature is not raised above 40° C. so that labile aqueous adjuvantsor labile delivered agents are not degraded. Preferably, once thedelivery system has coalesced, other components such as fragrances andcolors are added or the delivery system is incorporated into a cream orointment or a device for applying the delivery system.

Several formulations of delivery system are within the scope of aspectsof the invention. In embodiments that include an aqueous adjuvant, theratio of hydrophilic component:hydrophobic component:aqueous adjuvant isdesirably 3:4:3, but preferred formulations comprise 1:1:4, 1:1:14, and1:10:25. As described above, a sufficient amount of delivered agent tosuit the intended purpose is incorporated into the delivery system. Theamount of delivered agent that is incorporated into the penetrationenhancer depends on the compound, desired dosage, and application.

In some embodiments, the transdermal delivery composition is made byproviding an ethoxylated oil, mixing the ethoxylated oil with analcohol, non-ionic solubilizer, or emulsifier so as to form apenetration enhancer, mixing the penetration enhancer with an aqueousadjuvant (e.g., an extract from a plant of the Liliaeacae family), andmixing the penetration enhancer and aqueous adjuvant with a deliveredagent and thereby making the transdermal delivery composition. Forexample, an embodiment of a transdermal delivery composition comprisinga pain relief solution is manufactured as follows. A solution of 2.0% to7.0% oleoresin capsicum, 2.5 grams of Boswellin is mixed with 400 ml ofabsolute carpilic alcohol or isopropyl alcohol, 300 ml of ethoxylatedcastor oil, and 300 ml of a 100% solution of Aloe Vera. This transdermaldelivery composition has been observed to alleviate pain when rubbed ona targeted area.

The transdermal delivery compositions having a form of Hepsyl as adelivered agent desirably are comprised by weight or volume of between0.005% to 12.0% Hepsyl, depending on the type of Hepsyl, its solubility,and the intended application. For example, embodiments having Hepsyl CA1501C. Hepsyl CGA 1501K., and Hepsyl RA 150K can be comprised by weightor volume of 0.01-2 grams of Hepsyl delivered agent, 0-50 mL ofhydrophobic penetration enhancers (e.g., ethoxylated castor oil, jojobaoil, etc.), 0-50 mL of hydrophilic penetration enhancers, nonionicsolubilizers, or emulsifiers (e.g., isopropyl. alcohol, DMSO, etc.), and0-50 mL of aqueous adjuvant (e.g., water, Aloe Vera extract, etc.). Aparticularly desirable embodiment of the invention is comprised of0.1-0.5 gram of Hepsyl, 5-10 mL of ethoxylated castor oil, 5-10 mL ofisopropyl alcohol, and 5-10 mL of Aloe Vera extract. By using theseformulations, other delivered agents can be incorporated into atransdermal delivery composition. Formulations of transdermal deliverycompositions having collagens are described in the examples. Thefollowing section describes several therapeutic, prophylactic andcosmetic applications.

Therapeutic, Prophylactic, and Cosmetic Applications

Many embodiments are suitable for treatment of subjects either as apreventive measure (e.g., to avoid pain or skin disorders) or as atherapeutic to treat subjects already afflicted with skin disorders orwho are suffering pain. In general, most drugs, chemicals, and cosmeticagents that can be incorporated into a pharmaceutical or cosmetic can beformulated into a transdermal delivery composition of the invention.Because the various formulations of transdermal delivery compositiondescribed herein have a considerable range in hydrophobic andhydrophilic character, most drugs, chemicals, and cosmetic preparationscan be incorporated therein. That is, by adjusting the amount ofethoxylation, alcohol, and water in a particular formulation mostpharmaceutical and cosmetic agents are solubilized in a transdermaldelivery composition with little effort. Furthermore, because thetransdermal delivery compositions described herein can deliver a widerange of materials of both high and low molecular weight to skin cells,the utility of the transdermal delivery compositions described herein isincredibly broad. The aspects of the invention that follow are forexemplary purposes only, and one of skill in the art can readilyappreciate the wide spread applicability of a transdermal deliverycomposition described herein and the incorporation of other deliveredagents into a formulation of transdermal delivery composition isstraight forward.

In one embodiment, for example, a method of treatment or prevention ofinflammation, pain, or human diseases, such as cancer, arthritis, andAlzheimer's disease, comprises using a transdermal delivery compositiondescribed herein that has been formulated with an NSAID. Becausedelivered agents such as NSAIDs, capsaicin, and Boswellin interfereand/or inhibit cyclooxygenase enzymes (COX-1 and COX-2), they provide atherapeutically beneficial treatment for cancer and Alzheimer's diseasewhen administered by a transdermal delivery composition describedherein. (See U.S. Pat. No. 5,840,746 to Ducharme et al., and U.S. Pat.No. 5,861,268 to Tang et al.).

By one approach, a transdermal delivery composition comprising adelivered agent that is effective at reducing pain or inflammation(e.g., NSAIDS, capsaicin, Boswellin, or any combination thereof) isadministered to a subject in need and the reduction in pain orinflammation is monitored. An additional approach involves identifying asubject in need of a COX enzyme inhibitor (e.g., a subject sufferingfrom cancer or Alzheimer's disease) and administering a transdermaldelivery composition comprising a delivered agent that inhibits a COXenzyme (e.g., NSAIDS, capsaicin, Boswellin, or any combination thereof).Although many individuals can be at risk for contracting cancer orAlzheimer's disease, those with a family history or a genetic markerassociated with these maladies are preferably identified. Severaldiagnostic approaches to identify persons at risk of developing thesediseases have been reported. (See e.g., U.S. Pat. Nos., 5,891,857;5,744,368; 5,891,651; 5,837,853; and 5,571,671). The transdermaldelivery composition is preferably applied to the skin at a region ofinflammation or an area associated with pain or the particular conditionand treatment is continued for a sufficient time to reduce inflammation,pain, or inhibit the progress of the disease. Typically, pain andinflammation will be reduced in 5-20 minutes after application. Cancerand Alzheimer's disease can be inhibited or prevented with prolongeduse.

In another method, an approach to reduce wrinkles and increase skintightness and flexibility (collectively referred to as “restoring skintone”) is provided. Accordingly, a transdermal delivery compositioncomprising a form of collagen or fragment thereof as a delivered agentis provided and contacted with the skin of a subject in need oftreatment. By one approach, a subject in need of skin tone restorationis identified, a transdermal delivery composition comprising collagen ora fragment thereof is administered to the subject, and the restorationof the skin tone is monitored. Identification of a person in need ofskin restoration can be based solely on visible inspection and thedesire to have tight, smooth, and flexible skin. Treatment with thedelivery system is continued until a desired skin tone is achieved.Typically a change in skin tone will be visibly apparent in 15 days butprolonged use may be required to retain skin tightness and flexibility.The form of collagen in the delivered agent can be from various sourcesand can have many different molecular weights, as detailed above.Preferably, high molecular weight natural collagens are used, however,recombinant collagens, modified collagens, protease resistant collagens,and fragments thereof may be used with some of the transdermal deliverycompositions described herein.

The transdermal delivery compositions described herein can be processedin accordance with conventional pharmacological, veterinary andcosmetological methods to produce medicinal, veterinary, and cosmeticagents for administration to animals and humans in need thereof (e.g.,mammals including humans, dogs, cats, horses, cattle, and othercompanion or farm animals). The transdermal delivery compositionsdescribed herein can be incorporated into a pharmaceutical or cosmeticproduct with or without modification. The compositions of the inventioncan be employed in admixture with conventional excipients, e.g.,pharmaceutically acceptable organic or inorganic carrier substancessuitable for topical application that do not deleteriously react withthe molecules that assemble the delivery system. The preparations can besterilized and if desired mixed with auxiliary agents, e.g., lubricants,preservatives, stabilizers, coloring, aromatic substances and the likethat do not deleteriously react with the active compounds. They can alsobe combined where desired with other active agents. Embodimentsdescribed herein can be made according to good manufacturing processes(e.g., certified GMP), can be approved by a governmental body, such asthe Food and Drug Administration, and may have indicia that indicatesthat said compositions were manufactured GMP or were approved by agovernmental body, with or without structure-function indicia (e.g.,indicia that indicates the product's usefulness for improvement of one'sappearance or the general health and welfare of individuals that use theproduct).

The effective dose and method of administration of a transdermaldelivery system formulation can vary based on the individual patient andthe stage of the disease, as well as other factors known to those ofskill in the art. Although several doses of delivered agents have beenindicated above, the therapeutic efficacy and toxicity of such compoundsin a delivery system of the invention can be determined by standardpharmaceutical or cosmetological procedures with experimental animals,e.g., ED50 (the dose therapeutically effective in 50% of the population)and LD50 (the dose lethal to 50% of the population). The dose ratio oftoxic to therapeutic effects is the therapeutic index, and it can beexpressed as the ratio, LD50/ED50. Pharmaceutical and cosmetologicalcompositions that exhibit large therapeutic indices are preferred. Thedata obtained from animal studies is used in formulating a range ofdosages for human use. The dosage of such compounds lies preferablywithin a range of circulating concentrations that include the ED50 withlittle or no toxicity. The dosage varies within this range dependingupon the dosage form employed, sensitivity of the patient, and the routeof administration.

The exact dosage is chosen by the individual physician in view of thepatient to be treated. Dosage and administration are adjusted to providesufficient levels of the active moiety or to maintain the desiredeffect. Additional factors that may be taken into account include theseverity of the disease state, age, weight and gender of the patient;diet, time and frequency of administration, drug combination(s),reaction sensitivities, and tolerance/response to therapy. Short actingcompositions are administered daily whereas long acting pharmaceuticalcompositions are administered every 2, 3 to 4 days, every week, or onceevery two weeks. Depending on half-life and clearance rate of theparticular formulation, the pharmaceutical compositions of the inventionare administered once, twice, three, four, five, six, seven, eight,nine, ten or more times per day.

Routes of administration of the delivery systems of the invention areprimarily topical, although it is desired to administer some embodimentsto cells that reside in deep skin layers. Topical administration isaccomplished via a topically applied cream, gel, rinse, etc. containinga delivery system of the invention. Compositions of deliverysystem-containing compounds suitable for topical application include,but are not limited to, physiologically acceptable ointments, creams,rinses, and gels.

In some embodiments, the transdermal delivery composition isincorporated into a device that facilitates application. The embodiedcompositions generally have a vessel joined to an applicator, wherein atransdermal delivery composition of the invention is incorporated in thevessel. Some devices, for example, facilitate delivery by encouragingvaporization of the mixture. These apparatus have a transdermal deliverycomposition of the invention incorporated in a vessel that is joined toan applicator such as a sprayer (e.g., a pump-driven sprayer). Theseembodiments can also comprise a propellant for driving the incorporatedtransdermal delivery composition out of the vessel. Other apparatus canbe designed to allow for a more focused application. A device thatfacilitates a focused application of a transdermal delivery compositionof the invention can have a roll-on or swab-like applicator joined tothe vessel that houses the transdermal delivery composition. Severaldevices that facilitate the administration of a delivery system of theinvention have a wide range of cosmetic or therapeutic applications. Anexemplary transdermal delivery device is described in the section thatfollows.

Transdermal Delivery Dispenser

In some embodiments, the transdermal delivery composition is provided ina single dose application containing a pre-measured amount of adelivered agent. For example, septum sealed vials with or without anapplicator (e.g., a swab) containing a pre-measured amount oftransdermal delivery composition (e.g., 0.5 ml) containing apre-measured amount of a delivered agent (e.g., 400 mg of ibuprofen, 0.6mg marine collagen, or 1 g of aspirin) are embodiments of the invention.These embodiments have significant utility because pre-determined dosesof certain delivered agents facilitate appropriate treatment regimensand the individually sealed doses of the transdermal deliverycomposition with delivered agent maintain sterility of the compositionbetween applications.

FIGS. 1A and 1B show an exemplary embodiment of a dispenser 100. As canbe seen in FIG. 1A, in which the dispenser 100 is shown in an explodedstate, the dispenser 100 comprises a removable cartridge 102 and a bodyportion 104. A latch member 106 on the body portion 104, is shown in anunsecured state, permitting the insertion and removal of the removablecartridge 102. The latch member 106 is slidable between the unsecuredposition as shown and a secured position 108, shown in shadow, in whichthe insertion and/or removal of a removable cartridge 102 is inhibited.Although a slidable latch member 106 is shown in this embodiment, itwill be understood that any method of securing the removable cartridge102 to the body portion 104 can be used. For example, a pin attached tothe body portion 104 could engage an aperture on the removable cartridge102. Alternately, if the body portion is formed from a sufficientlyresilient material, the body portion can be designed such that a snugfit is formed without any need for additional securing methods.Transparent portion 103 permits the user to view the amount of fluidremaining in removable cartridge 102. Similarly, transparent portion 105allows the user to see the amount of fluid to be dosed.

FIG. 1B shows the dispenser 100 in an assembled state, where theremovable cartridge 102 has been inserted into the body portion 104. Thelatch member 106 has been moved to the secured position 108 shown inshadow in FIG. 1A.

FIG. 2 schematically depicts a cross-section of the dispenser 100 ofFIG. 1, in an assembled state. It can be seen that the removablecartridge 104 comprises a fluid reservoir 210, which is configured tohold the therapeutic drug delivery fluid. The removable cartridge 104further comprises a movable upper wall 212, which forms the upper wallof the fluid reservoir 210. The movable upper wall is displaceable in atleast the downward direction. The removable cartridge 102 also includesa one-way valve 214, such as a check valve, located at the bottom of theremovable cartridge, which is in fluid communication with the fluidreservoir 210. As will be described in greater detail below,displacement of the movable upper wall 212 in the downward directionwill cause fluid to flow from the fluid reservoir 210 through the valve214.

Still with respect to FIG. 2, the body portion 102 of the dispenser 100includes a dosing chamber 220. When the dispenser 100 is in an assembledspace, the dosing chamber 220 is in fluid communication with the valve214 of the removable cartridge via an aperture 222 in the dosing chamberaligned with the valve 214. The upper wall of the dosing chamber 220 isformed by the lower surface of a movable member 224, alternatelyreferred to as a dosing member. In this embodiment, movable member 224comprises a threaded aperture through which a threaded portion 232portion of shaft 230 extends. Stop members 234 a and 234 b are locatedat the upper and lower ends, respectively, of the shaft 230. Anon-threaded portion 236 of shaft 230 extends through an aperture in thetop of the body portion 104, and wide sections 238 a and 238 b of shaft230 constrain vertical translation of the shaft 230 with respect to thebody portion 102. A knob 239 at the top of the shaft 220 facilitatesrotation by a user.

The size of the dosing chamber 220 can be adjusted by rotating the knob239, causing rotation of the shaft 230. As the dosing chamber 220 andthe movable member 224 have a non-circular shape, the movable member 224cannot rotate along with the shaft 230. The rotational movement of theshaft therefore results in vertical translation of the movable member224, changing the volume of the dosing chamber 220. When the movablemember reaches one of stop members 234 a,b, the rotational movement ofthe shaft 230 will be inhibited. The movable member 224 may comprise aring of partially deformable material (not shown), such as a rubberizedmaterial, around the edges of the movable member which come in contactwith the walls of the dosing chamber, in order to facilitate a tightseal between the edges of the movable member and the walls of the dosingchamber, so as to prevent undesired leakage along the sides of themovable member.

The lower end of the dosing chamber 220 comprises a sloped surface 240,and an aperture 242 in the wall of the dosing chamber. This aperture 242preferably extends to the bottom of the dosing chamber at least onepoint along the bottom surface of the dosing chamber, such that allfluid in the dosing chamber 220 can flow out of the aperture 242.

The body portion 104 further comprises a plunger 250 having an upper end252 and a lower end 254 configured to engage the movable upper wall ofthe removable cartridge 102. The plunger 250 extends through an aperturein the top of the body portion 104. The plunger 250 is preferably biasedto return to a position in which the upper surface of the removablecartridge is not engaged. This may be done, for example, via a spring256 connecting the body portion 104 and the plunger 250. As will bediscussed in greater detail below, it may be desirable to permit theuser to control the timing of the return to the initial position. Thismay be accomplished via the inclusion of teeth 251 along the shaft ofthe plunger, and a locking member 257 which is attached to the interiorof the body portion 102 and biased to remain in a position against theplunger 250, as shown. The teeth 251 therefore permit the downwardmovement of the plunger, but inhibit the return of the plunger upward toits original position. Locking member 257 is operably connected to arelease button 258 on the exterior of the dispenser 100. Engaging therelease button rotates locking member 257 about pivot point 259, andpermits the return of the plunger to its original position. When therelease button is disengaged, the bias of locking member 257 returns itto the position shown in FIG. 2.

The body portion 104 further comprises a slidable member 260 which ismovable between a first position in which the slidable member 260inhibits fluid flow out of the dosing chamber 220 through the aperture242, and a second position in which the slidable member 250 inhibitsfluid flow from the removable cartridge 102 to the dosing chamber 220via aperture 222. The slidable member 260 is in the first position whenthe plunger is in a depressed position, and the second position when theplunger retracts to an undepresed position. This may be accomplished,for example, via spring 262, which connects the plunger 250 to theslidable member 262. When the plunger is depressed, the spring 262 holdsthe slidable member 260 within a slot 264, located below the slopedsurface 240 which forms the bottom of the chamber. As the pressureincreases, the sliding member is prevented from flexing away from thedosing chamber by tabs 266. Thus, when the sliding member 260 is in thefirst position, shown in FIG. 2, the dosing chamber can be filled andfluid will not leak out. Fluid is permitted to flow into the dosingchamber due to the shape of sliding member 260, discussed in greaterdetail with respect to FIG. 4. When the plunger is moved to anundepressed position, the slidable member will be pushed upward to thesecond position, where the flow of fluid through the aperture 242 ispermitted.

At the bottom of the dispenser 100 is an applicator. In the presentembodiment, the applicator consists of an ellipsoidal applicator 280mounted on pins 280 a and 280 b which extend at least partially into theapplicator 280 along the axis of the applicator. Applicator 280 thusprovides a roll-on applicator, such that once the therapeutic fluid isreleased from the dosing chamber after the release button 258 ispressed, the fluid will flow downwards onto applicator 280. Theapplicator can then be placed in contact with the skin of the user, andthe dispenser moved to cause the applicator to roll across the skin ofthe patient, applying the desired dose of the therapeutic fluid to thepatient. It will be understood that alternate non-invasive applicatorscan be used in place of the roll-on applicator. These alternatenon-invasive applicators may include, but are not limited to, anabsorbent applicator tip, such as a sponge, or an applicator surfacehaving perforations through which the therapeutic fluid can flow.

In the above embodiment, the dosing chamber will be partially filledwith fluid when the plunger is depressed, but the air in the dosingchamber will not be permitted to escape, and will therefore becompressed in the dosing chamber. In order to protect the dispenser 100from damage due to excessive pressure created in the dosing chamber, thevalve 214 may be designed to close when a certain pressure has beenreached. Taking into account this pressure, and the volume of thetrapped air at that pressure, accurate dosing can be obtained byaccounting for the volume of the trapped air in the dosing chamber.

In an alternative embodiment, the movable member 224 may comprise amechanism for allowing air to exit the dosing chamber without permittingfluid. An exemplary system for doing so is shown in FIGS. 3A and 3B. InFIG. 3A, it can be seen that the movable member 324 includes an aperture370. A sphere 372, which is buoyant relative to the fluid 376 which willbe used, is suspended within a track 374, which permits movement of thesphere 372 upward to engage the aperture 370, forming a seal, butinhibits movement of the sphere 372 downward below a level necessary toallows air to flow over the sphere 372 and out through the aperture 370.

FIG. 3B shows the dosing chamber full of fluid 376. The buoyant sphere372 is lifted as the fluid level rises within the dosing chamber.Because the sphere 372 is kept level with the fluid, almost all of theair is allowed to escape, but the fluid cannot escape through theaperture once the sphere engages the aperture. The sphere 372 mayadvantageously be formed of a partially deformable material, tofacilitate the forming of a seal between the sphere and the movablemember 324. Bouancy of the sphere 372 may be achieved through selectionof an appropriate material, or through the use of a hollow sphere, inorder to increase buoyancy.

It will be understood that alternate methods of permitting air to escapewhile preventing fluid flow through the movable member 220 may beutilized, including the use of specialized valves which permit the flowof air while inhibiting the flow of fluid through the valve.

The operation of the sliding member 370 is now described with respect toFIGS. 4A and 4B. FIG. 4A depicts a portion of the cross section of thedispenser 100 of FIG. 2, taken along line 4 of FIG. 2. In particular, itcan be seen that FIG. 4A depicts an embodiment in which the slidablemember 260, shown partially in shadow where it is locate behind otherfeatures, is in a first position in which the slidable member does notinhibit the flow of fluid from the fluid reservoir 210 through theaperture 222 into the dosing chamber 220 (for simplicity, the valve 214is not depicted, but would be in line with aperture 222). This is due tothe design of the slidable member 260 such that it is substantiallyL-shaped. It can be seen that when the slidable member 260 is in thisfirst position, flow of the fluid out of the dosing chamber 230 throughthe aperture 242 is prevented by the lower portion of slidable member260. Slidable member 260 is prevented from flexing away from aperture242 by the upper portions 266 a of tab members 266. In addition, thelower portions 266 b of tab members 266 prevent additional downwardmovement of slidable member 260. Thus, tab members 266 form a slot 264(see FIG. 2) which constrains the slidable member such that it forms asufficiently rigid barrier to constrain fluid flow out of the dosingchamber.

In FIG. 4B, the slidable member 260 has been moved to a second positionin which the slidable member 260 obstructs the flow of fluid throughaperture 222 into dosing chamber 220, but permits the flow of fluid fromdosing chamber 220 through aperture 242, and downward to applicator 280(not shown). Because the slidable member 260 is operably connected viaspring 262 to plunger 250, the slidable member 260 is in the firstposition when the plunger is depressed and fluid is being dispensed intothe dosing chamber, and in the second position when the plunger returnsto its original position after the release button is depressed. As canbe seen, the lower, thicker portion of the slidable member 260 desirablyhas sufficient height that at intermediate positions of the slidablemember, both of the apertures 222, 242 are completely occluded. Thus, noadditional fluid beyond what is already in the dosing chamber will bedispensed.

Although the above detailed description has shown, described, andpointed out novel features of the invention as applied to variousembodiments, it will be understood that various omissions,substitutions, and changes in the form and details of the deviceillustrated may be made by those skilled in the art without departingfrom the spirit of the invention.

For example, in another embodiment, the dispenser may not include afluid reservoir contained within a removable cartridge, but may insteadbe a disposable dispenser without a replaceable cartridge. In anotherembodiment, the volume of the dosing chamber need not be adjustable bythe user. Such an embodiment may be advantageous in situations whereprecise dosing is required, or where regular fixed doses are required.

In another embodiments, the user actuatable knob which controls the sizeof the dosing chamber need not be fixed directly to the rotatable shaft,but may instead be operably connected to the rotatable shaft via a gearor a series of gears, so as to facilitate either rapid adjustment of thedosing volume or very precise adjustment of the dosing volume, dependingon the relative properties of the gears. In alternate embodiments, oneor more of the operably connected features need not be mechanicallyconnected, as described and depicted above. For instance, electricalconnections between features and electrical actuators, such as servomotors, stepper motors, or hydraulics, can be used to replace themechanical interconnections described above. For example, the knob 239could be replaced by two buttons, electrically connected to a motor, oneof which causes the motor to drive the rotatable shaft in one direction,and the other of which causes the motor to drive the rotatable shaft inthe other direction. Similarly, the plunger could be replaced by aplunger which is electronically actuatable at the push of a button. Apressure sensor within the dosing chamber could be used to release theplunger once a sufficient pressure has been reached.

Thus, these and other modifications to the above described devices canbe made by persons having ordinary skill in the art without departingfrom the spirit of the invention. As will be recognized, the presentinvention may be embodied within a form that does not provide all of thefeatures and benefits set forth herein, as some features may be used orpracticed separately from others.

Example 1 below describes a clinical study that was performed toevaluate the efficacy of a transdermal delivery composition thatcomprised capsaicin.

EXAMPLE 1

In this example, evidence is provided that a transdermal deliverycomposition of the invention can administer a therapeutically effectiveamount of a low molecular weight delivered agent (e.g., 0.225% oleoresincapsicum). A clinical study was performed to evaluate the effectivenessof a transdermal delivery composition of the invention comprising 0.225%capsaicin (“EPRS”) as compared to a commercially available creamcomprising Boswellin, 10% methyl salicylate, and 0.25% capsaicin.(Nature's Herbs). The two pain relief preparations were tested on sixsubjects who suffer from degenerative arthritis, debilitating back pain,and/or bursitis. For the first five days of the study, the subjectsapplied the commercially available cream three times a day. On day six,application of the commercially available cream was stopped and subjectsapplied the EPRS transdermal delivery composition. The EPRS pain reliefsolution was also applied for five days, three times a day. Dailyanalysis of the efficacy of the particular pain relief formulations wastaken by the subjects and observations such as the time ofadministration, odor, and therapeutic benefit were recorded after eachadministration.

The five day use of the commercially available cream was found toprovide only minimal therapeutic benefit. The cream was reported toirritate the skin, have a noxious smell, and provide little decrease inpain or increase in flexibility or range of motion. In contrast, thefive day use of EPRS was reported to provide significant pain relief,relative to the relief obtained from the oral consumption of NSAIDs.Further, EPRS was reported to increase flexibility and range of motionwithin five to twenty minutes after application. Additionally, EPRS didnot present a significant odor nor did it cause skin irritation. Theresults of this study demonstrate that a delivery system comprising alow molecular weight compound, capsaicin, can effectively administer thedelivered agent to cells of the body where it provides therapeuticbenefit. The next example describes a clinical study that was performedto evaluate the efficacy of several different formulations oftransdermal delivery composition that comprised low and high molecularweight collagens.

EXAMPLE 2

In this example, evidence is provided that a transdermal deliverycomposition of the invention can administer a therapeutically effectiveamount of a low and high molecular weight delivered agent (e.g., a lowand high molecular weight collagens). A clinical study was performed toevaluate the effectiveness of several transdermal delivery compositionscomprising various penetration enhancers, aqueous adjuvants, andcollagen delivered agents. The various transdermal delivery compositionsthat were evaluated are provided in TABLE 18. Of the formulations thatwere originally screened, three were extensively evaluated by tensubjects (three men and seven women) in a single blind study. Theformulations analyzed in the single blind study are indicated in TABLE18 by a dagger. That is, the three different formulations (“P1”, “P2”,and “F4”) were evaluated.

The P1 formulation comprised approximately 0.73% to 1.46% Solu-Coll, asoluble collagen having a molecular weight of 300,000 daltons. The P2formulation comprised approximately 1.43% to 2.86% Plantsol, a plantcollagen obtained from yeast having a molecular weight of 500,000daltons. The F4 formulation comprised approximately 11.0% of HydroCollEN-55, a hydrolyzed collagen having a molecular weight of 2,000 daltons.The evaluation of the P1, P2, and F4 formulations was as follows. Left,right, and center mug-shot photographs were taken with a Pentax camerahaving a zoom 60× lens and Kodak-Gold 100 film before beginning thestudy. Shortly after, each subject was given a bottle having aformulation of transdermal delivery composition and was instructed toapply the solution to the right side of the face and neck, leaving theleft side untreated, twice daily for 15 days. The F4 formulation wastested first and the application was carried out after showering orwashing and before application of any other product to the treated areaof the face. After the 15 day period, three mug-shot photographs wereagain taken, the subjects recorded their observations on theeffectiveness of the formulation in a questionnaire, and a 7 day periodwithout application of a collagen product provided. The questionnairerequested the subject to assign a score (e.g., a numerical value thatrepresents effectiveness) on characteristics of the transdermal deliverycomposition formulation. Characteristics that were evaluated includedtackiness, odor, marketability, and overall effectiveness of theformulation, as well as, whether the formulation tightened the skin,decreased lines, conditioned or softened the skin, and had any negativeside-effects. The scale for the scoring was 1-10, with 1 being the worstrating and 10 being the best rating.

Following the test of F4, the evaluation detailed above was conducted onthe P1 formulation. Again, photographs were taken before and after thesecond 15 day protocol, a questionnaire evaluating the efficacy of theparticular formulation was completed, and a 7 day period withoutapplication of a collagen product was provided. Further, after the testof P1, the same evaluation was conducted on the P2 formulation,photographs were taken before and after the trial, and a questionnaireevaluating the efficacy of the particular formulation was completed.

The data from the three evaluation questionnaires were pooled, analyzedusing a “t-table” and standard deviation calculations were made. SeeTABLE 19. An overall rating for each particular formulation wasassigned. A perfect score by this system was a 7.875 overall rating. P1was found to have a 4.25 overall rating (approximately 54% effective),P2 was found to have a 4.625 overall rating (approximately 59%effective), and F4 was found to have a 5.625 overall rating(approximately 71% effective).

The before and after treatment photographs also revealed that the threetested transdermal delivery compositions provided therapeutic benefit. Adecrease in wrinkles was observed and an increase in skin tightness andfirmness can be seen. That is, P1, P2, and F4 all provided therapeuticand/or cosmetic benefit in that they restored skin tone in the subjectstested. The results presented above also demonstrate that transdermaldelivery compositions of the invention can be used to administer highmolecular weight delivered agents.

TABLE 18 ECO Aloe IPA Plantsol EN-55 Solu-coll DMPX YYO Score ID 29.7%*50.0%* 5.0%*  0* 8.3%*  0*  0*  0* 2 F-1 10.4% 79.0% 5.3% 0 8.7% 0 0 0 3F-2 5.2% 63.0% 5.3% 0 17.4% 0 0 0 3 F-3 5.0% 70.0% 5.0% 0 11.0% 0 0 0 3+F-4† 4.5% 18.2% 4.6% 0 0 0.7% to 1.5% 0 0 3+ P-1† 8.3% 8.3% 8.3% 0.7% to1.4% 4.6% 0.3% to 0.7% 0 0 2 Y-500 0.7% 22.2% 11.1% 1.3% to 2.7% 0 0 0 03+ P-501 0.4% 35.7% 3.6% 1.1% to 2.1% 0 0 0 0 2 P-502 0.9% 8.7% 0 0 02.3% to 4.6% 0 0 1 SC-1 1.8% 18.5% 0 0 44.8% 0 0 0 3+ SC-2 1.8% 17.9%7.1% 0 43.2% 0 0 0 3 SC-3 0.9% 9.4% 4.7% 0 34.3% 0.3% to 0.6% 0 0 1 PSCEN 1.8% 31.3% 6.3% 0.3% to 2.5% 0 0 0 0 3+ P-1A 0.8% 19.2% 3.8% 1.5% to3.1% 0 0 7.7% 0.3% 5 P-1C 0.7% 17.9% 7.1% 1.4% to 2.9% 0 0 1.1% 0.3% 5P-2† 0.7% 22.2% 11.1% 1.3% to 2.7% 0 0 0 0 3+ P-501 Abbreviations:ECO—ethoxylated castor oil (BASF) Aloe - Aloe Vera (Aloe Labs;(800)-258-5380) IPA - Absolute isopropyl alcohol (Orange CountyChemical, Santa Ana, California) Plantsol - Yeast extract collagen(Brooks Industries Inc., Code No. 06485) EN-55 - hydrolyzed bovinecollagen (Brooks Industries Inc., Code No. 01000) SoluColl—solublecollagen (Brooks Industries Inc., Code No. 01029) DMPX—dimethylpolysiloxane (5 centistokes) (Sigma) YYO—Y-ling-Y-lang oil (Young LivingEssential Oils, Lehl, Utah) ID—Identification number *The percentagesreflect volume to volume. †Sample used in the 45 day clinical trial.

TABLE 19 COLLAGEN T-TABLE Formulations standard P1 P2 F4 deviationTackiness 5 3 10 2.94 Skin tightness 7 5 8 1.25 Odor 2 8 8 2.83 Decreaselines 2 2 1 0.47 Soften skin 8 7 4 1.7 Total skin 5 5 6 0.47 restorationMarket Buying 5 7 8 1.25 Power Side effects 0 0 0 0 Total Score 4.254.63 5.63 1.36 (Average)

Several in vitro techniques are now widely used to assess thepercutaneous absorption of delivered agents. (See e.g., Bronaugh andCollier in In vitro Percutaneous absorption studies: Principle,Fundamentals, and Applications, eds. Bronaugh and Maibach, Boca Raton,Fl, CRC Press, pp 237-241 (1991) and Nelson et al., J. Invest. Dermatol.874-879 (1991)). Absorption rates, and skin metabolism can be studied inviable skin without the interference from systemic metabolic processes.The next example describes several approaches that can be used toevaluate the ability of a particular formulation of transdermal deliverycomposition to deliver a particular delivered agent.

EXAMPLE 3

Skin barrier function can be analyzed by examining the diffusion offluorescent and colored proteins and dextrans of various molecularweights (“markers”) across the skin of nude mice or swine. Swine skin ispreferred for many studies because it is inexpensive, can be maintainedat −20° C., and responds similarly to human skin. Prior to use, frozenswine skin is thawed, hair is removed, and subcutaneous adipose tissueis dissected away. Preferably, a thickness of skin that resembles thethickness of human skin is obtained so as to prepare a membrane thataccurately reflects the thickness of the barrier layer. A dermatome canbe pushed across the surface of the skin so as to remove any residualdermis and prepare a skin preparation that accurately reflects humanskin. Elevation of temperature can also be used to loosen the bondbetween the dermis and the epidermis of hairless skin. Accordingly, theexcised skin is placed on a hot plate or in heated water for 2 minutesat a temperature of approximately 50° C.-60° C. and the dermis isremoved by blunt dissection. Chemical approaches (e.g., 2M saltsolutions) have also been used to separate the dermis from the epidermisof young rodents.

Many different buffers or receptor fluids can be used to study thetransdermal delivery of delivered agents across excised skin prepared asdescribed above. Preferably, the buffer is isotonic, for example anormal saline solution or an isotonic buffered solution. Morephysiological buffers, which contain reagents that can be metabolized bythe skin, can also be used. (See e.g., Collier et al., Toxicol. Appl.Pharmacol. 99:522-533 (1989)).

Several different markers with molecular weight from 1,000 daltons to2,000,000 daltons are commercially available and can be used to analyzethe transdermal delivery compositions of the invention. For example,different colored protein markers having a wide range of molecularweights (6,500 to 205,000 daltons) and FITC conjugated protein markers(e.g., FITC conjugated markers from 6,500 to 205,000 daltons) areavailable from Sigma (C3437, M0163, G7279, A2065, A2190, C1311, T9416,L8151, and A2315). Further, high molecular weight FITC conjugateddextrans (e.g., 250,000, 500,000, and 2,000,000 daltons) are obtainablefrom Sigma. (FD250S, FD500S, and FD2000S).

Accordingly, in one approach, swine skin preparations, obtained asdescribed above, are treated with a delivery system lacking a deliveredagent and control swine skin preparations are treated with water.Subsequently, the skin is contacted with a 1 mM solution of a markerwith a known molecular weight suspended in Ringer's solution (pH 7.4) at37° C. After one hour, the skin is frozen and sliced at a thickness of 5μm. The sections are counter stained with 5 μg/ml propidium and, if themarker is FITC conjugated, the sections are analyzed by fluoresencemicroscopy. If the marker is a colored marker, diffusion of the markercan be determined by light microscope. The marker will be retained inthe upper layers of the stratum corneum in the skin(delete “untreatedmice”) but the skin treated with the delivery system will be found tohave the dye distributed throughout the stratum corneum and any dermallayer that remains.

Additionally, modifications of the experiments described above can beperformed by using a delivery system comprising various molecular weightmarkers. Accordingly, skin preparations are treated with the deliverysystem comprising one or more markers and control skin preparations aretreated with water. After one hour, the skin is frozen and sliced at athickness of 5 μm. The sections can be counter stained with 5 μg/mlpropidium iodide and can be analyzed by fluoresence microscopy (e.g.,when a fluorescent marker is used) or alternatively, the sections areanalyzed under a light microscope. The marker will be retained in theupper layers of the stratum corneum in the skin(delete “untreated mice”)but the skin treated with the delivery system will be found to have thedye distributed throughout the stratum corneum and any dermal layer thatremains.

In another method, the transdermal water loss (TEWL) of penetrationenhancer-treated skin preparations can be compared to that of untreatedskin preparations. Accordingly, skin preparations are obtained, asdescribed above, and are treated with a delivery system of the inventionlacking a delivered agent (e.g., a penetration enhancer). Control skinpreparations are untreated. To assess TEWL, an evaporimeter is used toanalyze the skin preparation. The Courage and Khazaka Tewameter TM210,an open chamber system with two humidity and temperature sensors, can beused to measure the water evaporation gradient at the surface of theskin. The parameters for calibrating the instrument and use of theinstrument is described in Barel and Clarys Skin Pharmacol. 8: 186-195(1995) and the manufacturer's instructions. In the controls, TEWL willbe low. In contrast, TEWL in penetration enhancer-treated skinpreparations will be significantly greater.

Further, skin barrier function can be analyzed by examining thepercutaneous absorption of labeled markers (e.g., radiolabeled,fluorescently labeled, or colored) across skin preparations in adiffusion chamber. Delivery systems of the invention having variousmolecular weight markers, for example, the proteins and dextransdescribed above, are administered to swine skin preparations. Swine skinpreparations are mounted in side-by-side diffusion chambers and areallowed to stabilize at 37° C. with various formulations of penetrationenhancer. Donor and receiver fluid volumes are 1.5 ml. After 1 hour ofincubation, a labeled marker is added to the epidermal donor fluid toyield a final concentration that reflects an amount that would beapplied to the skin in an embodiment of the invention. Five hundredmicroliters of receiver fluid is removed at various time points, anequal volume of penetration enhancer is added to the system. The aliquotof receiver fluid removed is then analyzed for the presence of thelabeled marker (e.g., fluorescent detection, spectroscopy, orscintillation counting). Control swine skin preparations areequilibrated in Ringer's solution (pH 7.4) at 37° C.; the sameconcentration of labeled marker as used in the experimental group isapplied to the donor fluid after one hour of equilibration; and 5001 ofreceiver fluid is analyzed for the presence of the marker. In theexperimental group, the steady-state flux of labeled marker in the skinwill be significantly greater than that of the control group. By usingthese approaches, several transdermal delivery compositions can beevaluated for their ability to transport low and high molecular weightdelivered agents across the skin. The next example describes severaldifferent formulations of transdermal delivery composition that weremade to comprise various delivered agents, demonstrating the wide-rangeof utility of aspects of the invention.

EXAMPLE 4

In this example, several different formulations of transdermal deliverycomposition containing various delivered agents are provided. Theformulations described include: compositions for removing age spots andrestoring skin brightness, compositions for advanced pain relief, musclerelaxers, hormone replacement products, wound healing formulations,products for reducing fine lines and wrinkles, stretch mark reducingproducts, growth factor products, and anti-psoriasis products.

SKIN BRIGHTENING OR AGE SPOT REDUCING PRODUCT: Melaslow (10%) 30 mlEthoxylated Macadamia nut oil 160 ml  (16 ethoxylations/molecule)Ethanol 80 ml Water 40 ml Marine collagen (1%) 40 ml Etioline (5%) 30 ml

This formulation was found to rapidly reduce the appearance of age spotsin a subject that applied daily amounts of the product for thirty days.

STRETCH MARK REDUCING PRODUCTS: FORMULATION #1 Eucalyptus oil 400 mlEthanol 180 ml Ethoxylated macadamia nut oil 180 ml (16ethoxylations/molecule) Distilled water 40 ml various perfumes wereadded including lemon oil or 30 drops lavender or 30 drops sweet orangeor 1 ml tangerine 30 drops FORMULATION #2 Eucalyptus oil 500 ml Ethanol225 ml Ethoxylated macadamia nut oil 225 ml (16 ethoxylations/molecule)Distilled water 50 ml FORMULATION #3 Eucalyptus oil (Kayuuputih oil) 400ml Ethanol 220 ml Ethoxylated macadamia nut oil 180 ml (16ethoxylations/molecule) Distilled water 40 ml Y-Ling-Y-Lang 22 dropsCoconut oil 3 ml

These formulations were found to rapidly reduce the appearance ofstretch marks in a subject that applied daily amounts of the productsfor thirty days.

TESTOSTERONE SUPPLEMENTATION PRODUCTS: FORMULATION #1 Ethanol 30 mlEthoxylated macadamia nut oil 30 ml (16 ethoxylations/molecule) Water 20ml Testosterone 10 ml (200 mg/ml) Coconut oil 10 drops FORMULATION #2Ethanol 40 ml Ethoxylated macadamia nut oil 40 ml (16ethoxylations/molecule) Water 5 ml Testosterone 5 ml (200 mg/ml) Coconutoil 10 drops Y-Ling-Y-Lang oil 10 drops FORMULATION #3 Testosterone 10ml (200 mg/ml) Ethanol 40 ml Ethoxylated macadamia nut oil 40 ml (16ethoxylations/molecule) Coconut oil 10 drops Y-Ling-Y-Lang oil 10 dropsWater 3 ml FORMULATION #4 Testosterone 1,000 mg in 5 ml Ethanol 50 mlEthoxylated macadamia nut oil 40 ml (16 ethoxylations/molecule) Water 5ml Y-Ling-Y-Lang oil 15 drops Rain water 15 drops

These formulations were found to rapidly increase the amount oftestosterone in the blood of a subject that applied approximately 0.5 mlof the product daily.

PAIN RELIEF PRODUCTS: FORMULATION #1 Ethyl alcohol 10.4 g White willowbark extract 10.4 g Glucosamine HCL 10 g MSM 10 g Chrondroitan sulfatesodium 10 g Marine collagen (1%) 100 ml Aloe Vera (whole leaf)concentrate 100 ml Ethoxylated macadamia nut oil 300 ml (16ethoxylations/molecule) Y-Ling-Y-Lang oil 28 drops Coconut oil 3 mlIbuprofen 16 g FORMULATION #2 Ibuprofen 3 g Methocarbanol 3 gChlorzoxazone 5 g Ethanol 75 ml Macadamia nut oil 75 ml (16ethoxylations/molecule) Aloe Vera (whole leaf) concentrate 5 mlY-Ling-Y-Lang oil 10 drops FORMULATION #3 Acetyl salicylic acid 22 gIbuprofen 8.5 g Ethanol (undenatured) 500 ml Ethoxylated macadamia nutoil 400 ml (16 ethoxylations/molecule) Distilled water 100 ml Peppermintoil 20 drops FORMULATION #4 Acetyl salicylic acid 44 g Undenaturedethanol 800 ml Ethoxylated macadamia nut oil 200 ml (16ethoxylations/molecule) Distilled water 40 drops Y-ling Y-lang oil 40drops Peppermint oil 40 drops FORMULATION #5 Acetyl salicylic acid 44 gUndenatured ethanol 900 ml Ethoxylated macadamia nut oil 1000 ml (16ethoxylations/molecule) Distilled water 100 ml Y-ling y-lang oil 40drops Peppermint oil 40 drops FORMULATION #6 Liquid aspirin 44 gUndenatured ethanol 800 ml Ethoxylated macadamia nut oil 200 ml (16ethoxylations/molecule) Distilled water 40 drops Y-ling y-lang oil 20drops Peppermint oil 40 drops Compounds brought into solution withslight heat.

These formulations were found to reduce pain in several subjects within5-20 minutes after application. Depending on the formulation, the periodof pain reduction lasted from 45 minutes (e.g., acetyl salicylic acidpreparations) to several hours (e.g., ibuprofen containingpreparations).

SKIN CARE/ANTI-PSORIASIS/ANTI-ECZEMA/WOUND HEALING PRODUCTS: FORMULATION#1 Dmae bitartrate 22.5 g Alpha lipoic acid 5 g Ethyl alcohol 25 mlMarine collagen (1%) 25 ml Aloe Vera 25 ml Macadamia nut oil (16ethoxylations/molecule)

The Dmae bitartrate and alpha lipoic acid was brought into solution andfiltered prior to mixture with the ethoxylated macadamia nut oil.

FORMULATION #2 Ichtyocollagene (1%) 500 ml Distilled water 248 ml LKEKK(SEQ. ID. No. 1) 1 vial (about 1 ml~10 μg) Ethoxylated macadamia nut oil150 ml (16 ethoxylations/molecule) Ethanol 25 ml Phenochem 39 ml (i.e.,a mixture of Methyl Paraben, Ethyl Paraben, Propyl Paraben, ButylParaben, and Isobutyl Paraben) FORMULATION #3 Distilled water 100 mlLKEKK (SEQ. ID. No. 1) 5 bottles (~50 μg) Ethoxylated macadamia nut oil40 ml (16 ethoxylations/molecule) Ethanol 5 ml

These formulations were found to improve the healing of a wound (alaceration) and were found to reduce psoriasis and eczema in anafflicted subject.

FORMULATION #4 Distilled Water with Sodium Bi Carbonate 18 ml (pH8.2-8.6) Hepsyl 5 g Ethoxylated macadamia nut oil 20 ml (16ethoxylations/molecule) Ethyl Alcohol Anhydrous 20 ml

This formulation reduces psoriasis and eczema in an afflicted subject.

PRODUCTS THAT REDUCE THE APPEARANCE OF FINE LINES AND WRINKLESFORMULATION #1 Ichtyocollagene (1%) 2,990 ml Distilled water 1,483 mlEthoxylated Macadamia nut oil 922 ml (16 ethoxylations/molecule) Ethanol150 ml Matrixyl (8%) 236 ml Phenochem 236 ml Ethoxydiglycol 33 mlFORMULATION #2 Ichtyocollagene (6%) 250 ml Distilled water 124 mlEthoxylated macadamia nut oil 78 ml (16 ethoxylations/molecule)Phenochem 20 ml Bio-ten 1 ml (Atrium Biotechnologies, Inc., Quebec,Canada) Ethanol 10 ml FORMULATION #3 Ichtyocollagene (1%) 500 mlDistilled water 250 ml Ethoxylated macadamia nut oil 125 ml (16ethoxylations/molecule) Ethanol 2 ml Bio-ten 3 ml Phenochem 40 mlFORMULATION #4 Ichtyocollagene (1%) 2,990 ml Distilled water 1,483 mlEthoxylated macadamia nut oil 922 ml (16 ethoxylations/molecule) Ethylalcohol 150 ml Matrixyl 236 ml Phenochem 236 ml FORMULATION #5Ichtyocollagene (1%) 1,994 ml Distilled water 999 ml Ethoxylatedmacadamia nut oil 675 ml (16 ethoxylations/molecule) Ethanol 100 mlBioserum 24 ml (Atrium Biotechnologies, Inc., Quebec, Canada) Phenochem157 ml FORMULATION #6 Ichtyocollagene (1%) 500 ml Distilled water 250 mlEthoxylated macadamia nut oil 168.75 ml (16 ethoxylations/molecule)Ethanol 25 ml Bioserum 10 ml Phenochem 43.75 ml FORMULATION #7Ichtyocollagene (1%) 1,000 ml Ethoxylated macadamia nut oil 338 ml (16ethoxylations/molecule) Distilled water 500 ml Ethanol 50 ml Matrixyl 76ml Phenochem 76 ml FORMULATION #8 Ichtyocollagene (1%) 22.55 mlDistilled Water 11.7 ml Ethoxylated macadamia nut oil 7 ml (16ethoxylations/molecule) Phenochem 0.5 ml Ethanol 1.5 ml Bio Serum 1 mlTOTAL 44.25 ml FORMULATION #9 Ichtyocollagene (1%) 15.03 ml DistilledWater 7.8 ml Ethoxylated macadamia nut oil 4.67 ml (16ethoxylations/molecule) Phenochem 0.333 ml Ethanol 1 ml Bio Serum 0.67ml TOTAL 29.5 ml FORMULATION #10 Ichtyocollagene (1%)(Marine 150 mlCollagen, Sderma) Distilled Water 400 ml Ethoxylated macadamia nut oil120 ml (16 ethoxylations/molecule) Ethyl Alcohol (anhydrous) 10 ml YlingYlang 16 drops Crodaderm B (Croda, Inc.) 5.0 ml Phenochem 2.0 ml Sepigel15 g

Formulation #10 is mixed in the order listed, and heated to 80° F.

These formulations were found to reduce the appearance of fine lines andwrinkles in subjects that applied the formulations daily for thirtydays. It should be noted that Bioserum, which is obtainable from AtriumBiosciences, Ontario Canada, may contain one or more of the following:placental protein, amniotic fluid, calf skin extract, and serum protein.Also, phenochem may contain one or more of the following: MethylParaben, Ethyl Paraben, Propyl Paraben, Butyl Paraben, and IsobutylParaben, and sodium methylparaban imidizolidinyl urea. Additionalcomponents that may be included in some formulations of products thatreduce the appearance of fine lines and wrinkles include: igepal cephenedistilled, synasol, ethoxylated glycerides, trisodium EDTA, potassiumsorbate, citric acid, ascorbic acid, and distilled water. For example,one formulation contains: Collagen (Marine), Distilled Water, IgepalCephene Distilled, Methyl Paraben, Ethyl Paraben, Propyl Paraben, ButylParaben, Isobutyl Paraben, Synasol, Serum Protein, Purified Water,Amniotic Fluid. Placental Protein. Calfskin Extract, Hydrolyzed CollagenSodium Methylparaben Imidazolidinyl Urea. Ethoxylated Glycerides,Trisodium EDTA, Potassium Sorbate, Citric Acid, and Ascorbic Acid.

SPOT FAT REDUCERS FORMULATION #1 Epigallocatechin Gallate (ECGC) 40 gEthyl Alcohol 100 ml Distilled Water 100 ml Ethoxylated macadamia nutoil 60 ml (16 ethoxylations/molecule) Lipase 1 ml FORMULATION #2Epigallocatechin Gallate 40 g (ECGC)(DSM, Netherlands) Ethyl Alcohol 100ml Distilled Water 100 ml Ethoxylated macadamia nut oil 60 ml (16ethoxylations/molecule) Lipase 1 ml Caffeine 2.0 g

These formulations were found to reduce fat when applied to the body inindividual. Polyphenols other than ECGC, such as analogs of green teaextract, are suitable in the above formulations and can be substitutedfor or used in combination with ECGC in the above formulations. Thefollowing example describes experiments that employed two different skincell model systems to evaluate the ability of a transdermal deliverycomposition containing collagen to transport collagen to skin cells.

EXAMPLE 5

In this example, it is shown that a transdermal delivery composition ofthe invention comprising marine type 1 collagen or native collagenefficiently transported the delivered agent to skin cells. Two differentin vitro skin cell model systems were used, human cadaver skin and acellulose acetate skin cell model system. Based on the physiology of theskin, three possible pathways exist for passive transport of moleculesthrough the skin to the vascular network: (1) intercellular diffusionthrough the lipid lamellae; (2) transcellular diffusion through both thekeratinocytes and lipid lamellae; and (3) diffusion through appendages(hair follicles and sweat ducts). The cellulose acetate skin modelevaluates the ability of the delivered agent to transport using thefirst two pathways and the human cadaver skin evaluates the ability touse all three pathways.

In brief, the transdermal delivery composition comprising collagen wasapplied to the cellulose acetate and the human cadaver skin in adiffusion chamber and the results were recorded after 10 minutes, 30minutes and one hour. The diffused material was analyzed by aspectrophotometer (Hitachi U2000 multiscan spectrophotometer). A portionof the diffused material was also separated on a gel by electrophoresisand the collagen was stained using a collagen-specific dye. A portion ofthe diffused material was also immunoprecipitated using polyclonalantibodies specific for collagens types 1-7 and the immunoprecipitateswere analyzed by immunodiffusion.

The table below provides the collagen concentration in the varioussamples of transdermal delivery compositions tested. The proteinconcentration was determined using a micro-protein assay (Bio-Rad).

TABLE 20 Protein Concentrations Sample number Native type 1 CollagenMarine type 1 collagen Sample 1 0.40 mg/ml 1.14 mg/ml Sample 2 0.44mg/ml 1.09 mg/ml Sample 3 0.42 mg/ml 1.14 mg/ml Average 0.42 1.12Standard error 0.011 0.017 Variance 0.0004 0.0008 Standard deviation0.02 0.03

Penetration Analysis

The transdermal delivery composition containing either marine collagenor native collagen was applied to the human cadaver skin and thecellulose acetate skin model systems. The penetration studies wereperformed in a diffusion chamber and the results were recorded at 10minutes, 30 minutes and an hour later. Sections of skin or celluloseacetate were stained with a collagen specific dye and a light microscopewas used to visualize the transported collagen. TABLE 21 provides theresults of these experiments. Note, that the native collagen appeared topenetrate the skin in less time than the marine collagen. This may bedue to the differing concentrations of collagen used in the transdermaldelivery compositions (i.e., the concentration of the native collagenwas 0.40 mg/ml and the concentration of the marine collagen was 1.14mg/ml). Nevertheless, by one hour, almost all of both types of collagenhad penetrated the skin in the model systems employed.

TABLE 21 Percent Penetration as per time interval 10 Product Hydrodermminutes 20 minutes 30 minutes 60 minutes Marine Collagen Vial A SampleA1 40% 60% 75% 95% Sample A2 40% 60% 75% 95% Sample A3 40% 60% 75% 95%Marine Collagen Vial B Sample B1 40% 60% 75% 95% Sample B1 40% 60% 75%95% Sample B1 40% 60% 75% 95% Marine collagen Vial C Sample C1 40% 60%75% 95% Sample C1 40% 60% 75% 95% Sample C1 40% 60% 75% 95% NativeCollagen Sample 1 80% 95% Sample 2 80% 95% Sample 3 80% 95%

When similar concentrations of native collagen and marine collagen wereused in a transdermal delivery composition, the native collagen and themarine collagen penetrated the upper three layers of the epidermis inapproximately one hour. The marine collagen and the native collagen werelocalized in the upper three layers of the human cadaver epidermis usinga collagen specific dye. A similar distribution of the collagen wasconfirmed by the cellulose acetate skin model. See TABLES 22 and 23.

TABLE 22 PENETRATION IN THE LAYERS OF THE HUMAN SKIN EPIDERMISPenetration of Epidermis layers of the Skin (Human Skin diffusionchamber study) Stratum Stratum Stratum Stratum Stratum Corneum lucidumGranulosum Spinosum Basale Marine collagen Vial A Sample A1 ✓ ✓ ✓ — —Sample A2 ✓ ✓ ✓ — — Sample A3 ✓ ✓ ✓ — — Marine collagen Vial B Sample B1✓ ✓ ✓ — — Sample B1 ✓ ✓ ✓ — — Sample B1 ✓ ✓ ✓ — — Marine collagen Vial CSample C1 ✓ ✓ ✓ — — Sample C1 ✓ ✓ ✓ — — Sample C1 ✓ ✓ ✓ — — Nativecollagen Sample 1 ✓ ✓ ✓ — — Sample 2 ✓ ✓ ✓ — — Sample 3 ✓ ✓ ✓ — — Note:(✓) indicates the presence of the product in the above layers of theepidermis as determined by collagen specific staining observed by lightmicroscopy after one hour of product application. (—) indicates absenceof products in these layers of the epidermis.

TABLE 23 Penetration Hydroderm in Epidermis layers of the Skin(Cellulose Acetate model skin diffusion chamber study) Stratum StratumStratum Stratum Stratum  Corneum lucidum Granulosum Spinosum BasaleMarine collagen Vial A Sample A1 ✓ ✓ ✓ — — Sample A2 ✓ ✓ ✓ — — Sample A3✓ ✓ ✓ — — Marine collagen Vial B Sample B1 ✓ ✓ ✓ — — Sample B1 ✓ ✓ ✓ — —Sample B1 ✓ ✓ ✓ — — Marine collagen Vial C Sample C1 ✓ ✓ ✓ — — Sample C1✓ ✓ ✓ — — Sample C1 ✓ ✓ ✓ — — Native Collagen Sample 1 ✓ ✓ ✓ — — Sample2 ✓ ✓ ✓ — — Sample 3 ✓ ✓ ✓ — — Note: (✓) indicates the presence of theproduct in the above layers of the epidermis as determined by collagenspecific staining observed by light microscopy after one hour of productapplication. (—) indicates absence of products in these layers of theepidermis.

Spectrophotometric Analysis

Spectrophotometric analysis of the diffused material revealed that thetransdermal delivery composition enabled significant transport of bothtypes of collagens. See TABLE 24.

TABLE 24 Spectral Absorbance at wavelength 280 nm Sample number Nativetype 1 collagen Marine type 1 collagen Sample 1 2.35 2.832 Sample 22.766 2.772 Sample 3 2.751 2.683 Average 2.622 2.762 Standard error0.136 0.043 Variance 0.0557 0.0056 Standard deviation 0.24 0.07

Electrophoresis Analysis

A portion of the diffused material was then separated by electrophoresisand visualized by staining with a collagen-specific dye. The penetratedmarine collagen remained intact during and after the analysis becausethe labeled marine collagen detected in the diffused material wasobserved to have the same molecular weight as marine collagen that hadnot undergone the analysis (control sample). The results showed that themarine collagen prior to the penetration study and after the penetrationstudy maintained its molecular structure around 500 kilodaltons (KD).The native collagen also maintained a molecular weight around 500 KDbefore and after penetration of the epidermis, demonstrating that thenative collagen that was delivered by the transdermal deliverycomposition, like the marine collagen, remained intact into theepidermis.

Immunoprecipitation Analysis

When the transdermal delivery composition containing marine collagen wasimmunoprecipitated using polyclonal antibodies specific for collagenstypes 1-7 before and after the penetration study, more evidence that themarine collagen remained in tact after the transdermal delivery wasobtained. Immuno-diffusion studies verified that the marine collagenprior to penetration of the skin and post penetration of skin consistedmainly of type I collagen. This further confirmed that the collagenremained intact post penetration.

The penetration study described above provided strong evidence that thetransdermal delivery compositions described herein are effective attransporting high molecular weight molecules to skin cells. It wasfound, for example, that marine collagen type 1 (˜500 kD) effectivelypenetrated the upper 3 layers of the epidermis and remained intactwithin an hour. These findings were supported by histology,spectrophotometric analysis, electrophoretic separation analysis,immunoprecipitation analysis, and immuno-diffusion analysis. Thefollowing example describes a clinical study that was performed, whichverified that the transdermal delivery compositions described hereineffectively reduce wrinkles and improve skin tone in humans in needthereof.

EXAMPLE 6

A clinical study was performed to evaluate the ability of a transdermaldelivery composition containing collagen, prepared as described herein,to reduce wrinkles and fine lines and otherwise restore skin tone tosubjects in need thereof. The medial half of the facial region includingthe neck and the upper chest areas were assigned as the regions underinvestigation. During a subject's routine application of the product,three times a day, digital pictures were taken at days 0, 3, 7, 14 and21 of the regions under investigation of the face including thesymmetrical region of the face where the product was not applied.Micrometer measurements of the wrinkles were then made from the digitalpictures and also from the facial areas under investigation.

Subjects invited to participate in the study had facial wrinkles andwere 25 years or older. Non-facial wrinkle individuals were also invitedand served as the control group. The source of subjects for the studywas randomly selected from the ethnically diverse population group agesranging from 25 years to 88 years old.

TABLE 25 Description of the subjects participating in the studyIdentification Number Gender Ethnicity Age General Description F101601Female Hispanic 88 Distinct facial wrinkles American F101602 FemaleHispanic 67 Distinct facial wrinkles American F101603 Female Hispanic 25Distinct facial wrinkles American around the eyes F101604 FemaleCaucasian 28 Distinct facial wrinkles around the eye region M101605 MaleAsian 40 Distinct facial wrinkles around the eye region

Subjects that signed the study consent form received 30 mls of atransdermal delivery composition comprising marine collagen. Micrometermeasurements of the wrinkles were performed using a 10× magnificationobjective eye piece. The measurements were recorded and tabulatedtogether with the digital photographs before and after application ofthe product. The wrinkle measurements were determined within the 3-weekduration of the study. The tabulated results provided in TABLE 26, whichindicates the general observations by subjects utilizing the product,and TABLE 27, which shows the wrinkle measurements. TABLE 28 shows theaverage percent of wrinkle reduction data generated after 21 days ofapplication of the transdermal delivery composition comprising collagen.

TABLE 26 Days of product application on one half of the face includingIdentification the upper chest and neck regions Number Day 3 Day 7 Day14 Day 21 F101601 Skin felt soft, and The right half of The right halfof The right half of clear, when the face cleared the face cleared theface cleared compared to the up and felt up and felt up and felt otherhalf without smooth, the slight smooth, the smooth, the slight productapplication, burning sensation slight burning burning sensation slightburning was still present sensation no no longer present. sensation for3-5 for 3-5 minutes. longer present. minutes upon product application.F101602 Skin felt soft, and The right half of The right half of Theright half of clear, when the face cleared the face cleared the facecleared compared to the up and felt up and felt up and felt other halfwithout smooth, the slight smooth, the smooth, the slight productapplication, burning sensation slight burning burning sensation slightburning was still present sensation no no longer present. sensation for3-5 for 3-5 minutes. longer present. minutes upon product application.F101603 Skin felt soft, and The right half of The right half of Theright half of clear, when the face cleared the face cleared the facecleared compared to the up and felt up and felt up and felt other halfwithout smooth, the slight smooth, the smooth, the slight productapplication, burning sensation slight burning burning sensation slightburning was still present sensation no no longer sensation for 3-5 for3-5 minutes. longer present. present.. minutes upon product application.F101604 Skin felt soft, and The skin felt Developed The rashes clear,when smooth and very rashes in the cleared up, and compared to the softin the facial neck region, the skin had other half without region wherestopped using normal product application, product was product.appearance as the slight burning applied. other half in sensation for3-5 which the minutes upon product was not product application. applied.M101605 Skin felt soft, and The right half of The right half of Theright half of clear, when the face cleared the face cleared the facecleared compared to the up and felt up and felt up and felt other halfwithout smooth, the slight smooth, the smooth, the slight productapplication, burning sensation slight burning burning sensation slightburning was still present sensation still still present for 3-5sensation for 3-5 for 3-5 minutes. present for 3-5 minutes. minutes uponminutes. product application.

TABLE 27 Average wrinkle measurements with product application on onehalf of the face including the upper chest and neck areas in μmSubject's Regions Identification of the Number face Day 0 Day 3 Day 5Day 7 Day 14 Day 21 F101601 Around eyes   6 μm   6 μm   6 μm   5 μm 4.5μm 4.5 μm Temporal   7 μm   7 μm   7 μm   7 μm   6 μm 5.5 μm cheek Chin7.5 μm 7.5 μm 7.5 μm 7.5 μm 7.0 μm 6.5 μm Around 6.5 μm 6.5 μm 6.5 μm6.5 μm 6.0 μm 5.5 μm mouth F101602 Around eyes 3.5 μm 3.5 μm 3.5 μm 3.5μm 3.5 μm 3.2 μm Temporal 4.1 μm 4.1 μm 4.1 μm 4.1 μm 3.9 μm 3.5 μmcheek Chin 2.5 μm 2.5 μm 2.5 μm 2.5 μm 2.0 μm 2.0 μm Around 2.0 μm 2.0μm 2.0 μm 2.0 μm 2.0 μm 2.0 μm mouth F101603 Around eyes 1.5 μm 1.5 μm1.5 μm 1.5 μm 1.5 μm 1.2 μm Temporal 1.0 μm 1.0 μm 1.0 μm 1.0 μm 1.0 μm1.0 μm cheek Chin 0.9 μm 0.9 μm 0.9 μm 0.9 μm 0.9 μm 0.85 μm  Around 0.5μm 0.5 μm 0.5 μm 0.5 μm 0.5 μm 0.45 μm  mouth F101604 Around eyes 0.2 μm0.2 μm 0.2 μm 0.2 μm 0.2 μm ** Temporal 1.5 μm 1.5 μm 1.5 μm 1.5 μm 1.5μm ** cheek Chin 1.0 μm 1.0 μm 1.0 μm 1.0 μm 1.0 μm ** Around 0.5 μm 0.5μm 0.5 μm 0.5 μm 0.5 μm ** mouth M101605 Around eyes 1.5 μm 1.5 μm 1.5μm 1.5 μm 1.5 μm 1.0 μm Temporal 0.5 μm 0.5 μm 0.5 μm 0.5 μm 0.5 μm 0.3μm cheek Chin 1.0 μm 1.0 μm 1.0 μm 1.0 μm 1.0 μm 0.9 μm Around 1.5 μm1.5 μm 1.5 μm 1.5 μm 1.5 μm 1.2 μm mouth Note ** Indicates the subjectstopped using the product.

TABLE 28 The percent reduction of wrinkle measurement on the regionsSubject's of the face at day 21 of Identification Hydroderm productapplication Number Around eyes Temporal cheek Chin Around mouth F101601  25% 21.4% 13.3% 15.4% F101602  8.6% 14.6% 20.0%  0.0% F101603 20.0% 0.0%  5.6% 10.0% F101604  0.0%  0.0%  0.0%  0.0% M101605 33.3% 40.0%  10% 20.0% Average % 17.42%  15.20%  9.78% 9.08% Overall On the entirefacial region where the 10.29%  effectiveness product was applied.

The data generated from this study indicates that the overalleffectiveness of transdermal delivery composition comprising marinecollagen as a wrinkle reducer is 10.29% when applied twice daily for 21days. As indicated by Table 28, the percent reduction of the wrinklesvaries with the various areas of the face where it is applied, with17.4% reduction around the eye regions and 15.20% at the temporal cheeksat the high end and around 9% at the chin and mouth regions. The nextexample sets forth experiments that demonstrate that transdermaldelivery compositions containing ethoxylated oils of less than 20ethoxylations/molecule transfer a delivered agent to the skin moreeffectively than transdermal delivery compositions containingethoxylated oils of 20 or more ethoxylations/molecule.

EXAMPLE 7

Several transdermal delivery composition formulations containingcollagen (1.2 mg/ml) and an ethoxylated oil having different amounts ofethoxylations/molecule are prepared. Formulations containing ethoxylatedoil of either 12, 16, 18, 20, 24, and 30 ethoxylations/molecule, water,and marine collagen (1.2 mg/ml) are made. Approximately 0.5 ml of eachof these formulations are applied to human cadaver skin in a diffusionchamber and the penetration of collagen is monitored over time (e.g., 10minutes, 30 minutes, 45 minutes and one hour). Sections of the skin aretaken, stained with a collagen specific dye, and the stained sectionsare analyzed under a light microscope.

A greater amount of collagen-specific staining will be seen in stainedskin sections collected at the various time points with formulationscontaining less than 20 ethoxylations/molecule than with formulationscontaining 20 or more ethoxylations/molecule. Formulations containingless than 20 ethoxylations/molecule will also penetrate the skin fasterthan formulations containing 20 or more ethoxylations/molecule.

In a second set of experiments, the collagen that has penetrated theskin at the various time points above is collected from the diffusionchamber and analyzed in a spectrophotometer. As above, a greater amountof collagen will be detected in samples collected at the various timepoints with formulations containing less than 20 ethoxylations/moleculethan formulations containing 20 or more ethoxylations/molecule.Formulations containing less than 20 ethoxylations/molecule will also beobserved to penetrate the skin faster than formulations containing 20 ormore ethoxylations/molecule. The next example sets forth experimentsthat demonstrate that transdermal delivery compositions containingethoxylated fatty acids having 10-19 ethoxylations/molecule transfer adelivered agent to the skin as effectively as transdermal deliverycompositions containing ethoxylated oils having 10-19ethoxylations/molecule.

EXAMPLE 8

A transdermal delivery composition containing collagen (1.2 mg/ml) andan ethoxylated fatty moiety having 16 ethoxylations/molecule, water andmarine collagen is made. Several transdermal delivery compositionscontaining ethoxylated fatty moieties and having 16ethoxylations/molecule, water, and marine collagen are made.Approximately 0.5 ml of each of these formulations are applied to humancadaver skin in a diffusion chamber and the penetration of collagen ismonitored over time (e.g., 10 minutes, 30 minutes, 45 minutes and onehour). Sections of the skin are taken, stained with a collagen specificdye, and the stained sections are analyzed under a light microscope.

The same amount of collagen-specific staining will be seen in stainedskin sections collected at the various time points with formulationscontaining ethoxylated fatty moieties as compared to formulationscontaining ethoxylated oils. Formulations containing ethoxylated fattymoieties will also penetrate the skin at approximately the same rate ascompared to formulations containing ethoxylated oils.

In a second set of experiments, the collagen that has penetrated theskin at the various time points above is collected from the diffusionchamber and analyzed in a spectrophotometer. As above, approximately thesame amount of collagen will be detected in samples collected at thevarious time points with formulations containing ethoxylated fattymoieties as compared to formulations containing ethoxylated oils.Formulations containing ethoxylated fatty moieties will also be observedto penetrate the skin at approximately the same rate as formulationscontaining ethoxylated oils.

EXAMPLE 9

A transdermal delivery composition containing collagen (1.2 mg/ml) andan ethoxylated oil having 16 ethoxylations/molecule, water and marinecollagen is made. A portion of the composition is transferred to acartridge adapted for the exemplary transdermal delivery devicedescribed herein. The transdermal delivery device is preset to loadapproximately 0.5 ml of the formulation. Approximately 0.5 ml of theformulation is applied to human cadaver skin, either manually, or usingthe transdermal delivery device, in a diffusion chamber and thepenetration of collagen is monitored over time (e.g., 10 minutes, 30minutes, 45 minutes and one hour). Sections of the skin are taken,stained with a collagen specific dye, and the stained sections areanalyzed under a light microscope. Several samples are prepared andtreated at the same time.

The amount of collagen-specific staining seen in stained skin sectionscollected is substantially more consistent in the samples in which theformulation is administered via the transdermal delivery device than inthe samples in which the formulation is delivered manually.

In a second set of experiments, the collagen that has penetrated theskin at the various time points above is collected from the diffusionchamber and analyzed in a spectrophotometer. As above, the amount ofcollagen detected in samples collected from the various samples in whichthe formulation is delivered via the transdermal delivery device showsconsiderably less variation than the amounts of collagen calculated fromsamples in which the formulation was applied manually.

Although the invention has been described with reference to embodimentsand examples, it should be understood that various modifications can bemade without departing from the spirit of the invention. Accordingly,the invention is limited only by the following claims.

1. A transdermal delivery composition comprising an ethoxylated fattymoiety or lipid moiety and a delivered agent, wherein the amount ofethoxylation of said fatty moiety or lipid moiety is 10, 11, 12, 13, 14,15, 16, 17, 18, or 19 ethoxylations per molecule, and wherein said fattymoiety or lipid moiety comprises 10 carbon residues.
 2. The transdermaldelivery composition of claim 1 further comprising water.
 3. Thetransdermal delivery composition of claim 1 further comprising alcohol.4. The transdermal delivery composition of claim 1, wherein said fattymoiety or lipid moiety comprises at least 10 carbon residues but lessthan or equal to 30 carbon residues.
 5. The transdermal deliverycomposition of claim 1, wherein the amount of ethoxylation is the sameas the number of carbons residues in the fatty moiety or lipid moeity.6. The transdermal delivery composition of claim 1, wherein the fattymoiety or lipid moiety chain length is 10 carbon residues in length. 7.The transdermal delivery composition of claim 1, wherein the fattymoiety chain length is 12 carbon residues in length.
 8. The transdermaldelivery composition of claim 1, wherein the fatty moiety chain lengthis 14 carbon residues in length.
 9. The transdermal delivery compositionof claim 1, wherein the fatty moiety chain length is 16 carbon residuesin length.
 10. The transdermal delivery composition of claim 1, whereinthe fatty moiety chain length is 18 carbon residues in length.
 11. Thetransdermal delivery composition of claim 1, wherein the fatty moietychain length is 20 carbon residues in length.
 12. The transdermaldelivery composition of claim 1, wherein the fatty moiety chain lengthis 22 carbon residues in length.
 13. The transdermal deliverycomposition of claim 1, wherein the fatty moiety chain length is 24carbon residues in length.
 14. The transdermal delivery composition ofclaim 1, wherein the fatty moiety is a fatty acid.
 15. The transdermaldelivery composition of claim 14, wherein the fatty acid is unsaturated.16. The transdermal delivery composition of claim 14, wherein the fattyacid is polyunsaturated.
 17. The transdermal delivery composition ofclaim 14, wherein the fatty acid is palmitoleic acid (C16:1, delta 9).18. The transdermal delivery composition of claim 14, wherein the fattyacid is selected from the group consisting of C16:1, delta 1; C16:1,delta 2; C16:1, delta 3; C16:1, delta 4; C16:1, delta 5; C16:1, delta 6;C16:1, delta 7; C16:1, delta 8; C16:1, delta 9; C16:1, delta 10; C16:1,delta 11; C16:1, delta 12; C16:1, delta 13; and C16:1, delta
 14. 19. Thetransdermal delivery composition of claim 1, wherein said fatty moietyis a fatty amine.
 20. The transdermal delivery composition of claim 1,wherein said lipid moiety is a sphingolipid.
 21. The transdermaldelivery composition of claim 1, wherein said lipid moiety is aglycolipid.
 22. The transdermal delivery composition of claim 1, whereinsaid lipid moiety is a glycosphingolipid.
 23. The transdermal deliverycomposition of claim 1, wherein said transdermal delivery systemcomprises a homogeneous mixture of an ethoxylated fatty moiety.
 24. Thetransdermal delivery composition of claim 1, wherein said transdermaldelivery system comprises a heterogeneous mixture of ethoxylated fattyacids.
 25. The transdermal delivery composition of claim 1, wherein thedelivered agent is a hormone.
 26. The transdermal delivery compositionof claim 25, wherein said hormone is a polypeptide.
 27. The transdermaldelivery composition of claim 26, wherein the polypeptide is selectedfrom the group consisting of oxytocin, vasopressin,melanocyte-stimulating hormone, corticortropin, lipotropin, thyrotropin,growth hormone, prolactin, luteinizing hormone, human chorionicgonadotropin, follicle stimulating hormone, corticotropin-releasingfactor, gonadotropin-releasing factor, prolactin-releasing factor,prolactin-inhibiting factor, growth-hormone releasing factor,somatostatin, thyrotropin-releasing factor, calcitonin, calcitoningene-related peptide, parathyroid hormone, glucagon-like peptide 1,glucose-dependent insulinotropic polypeptide, gastrin, secretin,cholecystokinin, motilin, vasoactive intestinal peptide, substance P,pancreatic polypeptide, peptide tyrosine tyrosine, neuropeptidetyrosine, amphiregulin, insulin, glucagon, placental lactogen, relaxin,angiotensin II, atrial natriuretic peptide, and melatonin.
 28. Thetransdermal delivery composition of claim 25, wherein the hormone is anon-peptide hormone.
 29. The transdermal delivery composition of claim25, wherein the hormone is selected from the group consisting ofthyroxine, triiodothyronine, estradiol, estrone, progesterone,testosterone, cortisol, corticosterone, aldosterone, epinephrine,norepinepherine, and calctriol.
 30. The transdermal delivery compositionof claim 1, wherein said delivered agent is collagen.
 31. Thetransdermal delivery composition of claim 1, wherein said deliveredagent is an anesthetic.
 32. The transdermal delivery composition ofclaim 31, wherein said anesthetic is selected from the group consistingof articaine, procaine, tetracaine, chloroprocaine and benzocaine,novocain, mepivicaine, bupivicaine, benzocaine, and lidocaine.
 33. Thetransdermal delivery composition of claim 1, wherein said deliveredagent is an analgesic.
 34. The transdermal delivery composition of claim33, wherein said delivered agent is selected from the group consistingof tramadol hydrochloride, fentanyl, metamizole, morphine sulphate,ketorolac tromethamine, hydrocodone, oxycodone, morprhine, loxoprofen,Capsaicin, and Boswellin.
 35. The transdermal delivery composition ofclaim 1, wherein said delivered agent is an NSAID.
 36. The transdermaldelivery composition of claim 35, wherein said non-steroidal antiinflammatory drug (NSAID) is selected from the group consisting ofibuprofen (2-(isobutylphenyl)-propionic acid); methotrexate (N-[4-(2, 4diamino 6-pteridinyl-methyl]methylamino]benzoyl)-L-glutamic acid);aspirin (acetylsalicylic acid); salicylic acid; diphenhydramine(2-(diphenylmethoxy)-N,N-dimethylethylamine hydrochloride); naproxen(2-naphthaleneacetic acid, 6-methoxy-9-methyl-, sodium salt, (−));phenylbutazone (4-butyl-1,2-diphenyl-3,5-pyrazolidinedione);sulindac-(2)-5-fluoro-2-methyl-1-[[p-(methylsulfinyl)phenyl]methylene-]-1H-indene-3-aceticacid; diflunisal (2′,4′,-difluoro-4-hydroxy-3-biphenylcarboxylic acid;piroxicam(4-hydroxy-2-methyl-N-2-pyridinyl-2H-1,2-benzothiazine-2-carboxamide1,1-dioxide, an oxicam; indomethacin(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-H-indole-3-acetic acid);meclofenamate sodium (N-(2,6-dichloro-m-tolyl) anthranilic acid, sodiumsalt, monohydrate); ketoprofen (2-(3-benzoylphenyl)-propionic acid;tolmetin sodium (sodium 1-methyl-5-(4-methylbenzoyl-1H-pyrrole-2-acetatedihydrate); diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneaticacid, monosodium salt); hydroxychloroquine sulphate(2-{[4-[(7-chloro-4-quinolyl)amino]pentyl]ethylamino}ethanol sulfate(1:1); penicillamine (3-mercapto-D-valine); flurbiprofen([1,1-biphenyl]-4-acetic acid, 2-fluoro-alphamethyl-, (+−)); cetodolac(1-8-diethyl-13,4,9, tetrahydropyrano-[3-4-13]indole-1-acetic acid;mefenamic acid (N-(2,3-xylyl)anthranilic acid; and diphenhydraminehydrochloride (2-diphenyl methoxy-N,N-di-methylethamine hydrochloride).37. The transdermal delivery composition of claim 1, wherein thedelivered agent is an anti-inflammatory compound selected from the groupconsisting of hydrocortisone, prednisolone, triamcinolone, cortisone andpiroxicam.
 38. The transdermal delivery composition of claim 1, whereinthe delivered agent is an anti-infective compound selected from thegroup consisting of amoxicillin, clavulanate potassium, itraconazole,acyclovir, fluconazole, terbinafine hydrochloride, erythromycinethylsuccinate, acetyl sulfisoxazole, penicillin V, cephalexin,erythromycin, azithromycin, tetracycline, ciproflaxin, gentamycin,sulfathiazole, nitrofurantoin, norfloxacin, flumequine, and ibafloxacin,metronidazole, nystatin, lamivudine, indinavir sulfate, and stavudine.39. The transdermal delivery composition of claim 1, wherein thedelivered agent is metformin hydrochloride or acarbose.
 40. Thetransdermal delivery composition of claim 1, wherein the delivered agentis an immunogen.
 41. The transdermal delivery composition of claim 40,wherein the immunogen comprises a polynucleotide.
 42. The transdermaldelivery composition of claim 1 wherein the delivered agent is an immuneresponse modifier.
 43. The transdermal delivery composition of claim 42,wherein the immune response modifier is selected from the groupconsisting of purine derivatives, adenine derivatives, cyclosporine, andCpGs.
 44. The transdermal delivery composition of claim 1, wherein thedelivered agent is an enzyme inhibitor selected from the groupconsisting of zileuton, captopril, and lisinopril.
 45. The transdermaldelivery composition of claim 1, wherein the delivered agent is selectedfrom the group consisting of ergotamine, melatonin, sumatriptan,zolmitriptan, and rizatriptan.
 46. The transdermal delivery compositionof claim 1, wherein the delivered agent is selected from the groupconsisting of zolpidem, zolpidem tartrate, triazolam, and hycosinebutylbromide.
 47. The transdermal delivery composition of claim 1,wherein the delivered agent is selected from the group consisting ofiohexol, technetium, TC99M, sestamibi, iomeprol, gadodiamide, ioversol,and iopromide.
 48. The transdermal delivery composition of claim 1,wherein the delivered agent is selected from the group consisting ofalsactide, americium, betazole, histamine, mannitol, metyrapone,petagastrin, phentolamine, radioactive B₁₂, gadodiamide, gadopenteticacid, gadoteridol, perflubron, cyclosporine, sildenafil citrate,paclitaxel, ritonavir, and saquinavir.
 49. A transdermal deliverycomposition comprising a transdermal delivery enhancer comprising amultifunctional backbone, wherein said multifunctional backbonecomprises at least two reactive groups (R), wherein at least onereactive group is substituted with a fatty moiety, wherein at least onereactive group is substituted with a polyethoxy moiety, wherein R isselected from the group consisting of —OH, COOH, SH, and NH₂, whereinsaid fatty moiety is selected from the group consisting of a fatty acid,a fatty alcohol, a fatty amine, and a modified fatty acid; and adelivered agent.
 50. The transdermal delivery composition of claim 49,wherein said (R) groups on said multifunctional backbone arehomogeneous.
 51. The transdermal delivery composition of claim 49,wherein said (R) groups on said multifunctional backbone areheterogeneous.
 52. The transdermal delivery composition of claim 49,wherein said penetration enhancer comprises at least 10 ethoxylationsper molecule but less than or equal to 19 ethoxylations per molecule.53. The transdermal delivery composition of claim 49, wherein saidmultifunctional backbone is a tri-alcohol.
 54. The transdermal deliverycomposition of claim 53, wherein said fatty moiety is a fatty acid, andwherein said fatty acid comprises at least 10 carbon residues.
 55. Thetransdermal delivery composition of claim 49, wherein saidmultifunctional backbone is a triacid.
 56. The transdermal deliverycomposition of claim 55, wherein at least one of said (R) groups on saidtriacid is substituted with a fatty alcohol.
 57. The transdermaldelivery composition of claim 55, wherein said triacid has a benzenering.
 58. The transdermal delivery composition of claim 57, wherein saidtriacid is selected from the group consisting of hemi-mellitic acid,trimellitic acid, trimesic acid, and nitrilotriacetic acid.
 59. Thetransdermal delivery composition of claim 57, wherein said triacidcomprises a toluene group.
 60. The transdermal delivery composition ofclaim 57, wherein said triacid comprises a naphthalene group.
 61. Thetransdermal delivery composition of claim 49, wherein saidmultifunctional backbone is an amino acid.
 62. The transdermal deliverycomposition of claim 61, wherein said amino acid is selected from thegroup consisting of glutamic acid, aspartic acid, cysteine, glutamine,serine, threonine, tyrosine, lysine and aminomalonic acid.
 63. Thetransdermal delivery composition of claim 49, wherein saidmultifunctional backbone is triethanolamine.
 64. The transdermaldelivery composition of claim 49, wherein said multifunctional backboneis diethanolamine.
 65. The transdermal delivery composition of claim 49,wherein said multifunctional backbone is dimethylolurea.
 66. Thetransdermal delivery composition of claim 49, wherein saidmultifunctional backbone is glucosamine.
 67. The transdermal deliverycomposition of claim 49, wherein said multifunctional backbone is asugar.
 68. The transdermal delivery composition of claim 67, whereinsaid sugar comprises between 4 and 6 R groups, wherein said R groups are—OH.
 69. The transdermal delivery composition of claim 49, wherein thedelivered agent is a hormone.
 70. The transdermal delivery compositionof claim 69, wherein said hormone is a polypeptide.
 71. The transdermaldelivery composition of claim 69, wherein the polypeptide is selectedfrom the group consisting of oxytocin, vasopressin,melanocyte-stimulating hormone, corticortropin, lipotropin, thyrotropin,growth hormone, prolactin, luteinizing hormone, human chorionicgonadotropin, follicle stimulating hormone, corticotropin-releasingfactor, gonadotropin-releasing factor, prolactin-releasing factor,prolactin-inhibiting factor, growth-hormone releasing factor,somatostatin, thyrotropin-releasing factor, calcitonin, calcitoningene-related peptide, parathyroid hormone, glucagon-like peptide 1,glucose-dependent insulinotropic polypeptide, gastrin, secretin,cholecystokinin, motilin, vasoactive intestinal peptide, substance P,pancreatic polypeptide, peptide tyrosine tyrosine, neuropeptidetyrosine, amphiregulin, insulin, glucagon, placental lactogen, relaxin,angiotensin II, atrial natriuretic peptide, and melatonin.
 72. Thetransdermal delivery composition of claim 69, wherein the hormone is anon-peptide hormone.
 73. The transdermal delivery composition of claim72, wherein the hormone is selected from the group consisting ofthyroxine, triiodothyronine, estradiol, estrone, progesterone,testosterone, cortisol, corticosterone, aldosterone, epinephrine,norepinepherine, and calctriol.
 74. The transdermal delivery compositionof claim 49, wherein said delivered agent is collagen.
 75. Thetransdermal delivery composition of claim 49, wherein said deliveredagent is an anesthetic.
 76. The transdermal delivery composition ofclaim 75, wherein said anesthetic is selected from the group consistingof articaine, procaine, tetracaine, chloroprocaine and benzocaine,novocain, mepivicaine, bupivicaine, benzocaine, and lidocaine.
 77. Thetransdermal delivery composition of claim 49, wherein said deliveredagent is an analgesic.
 78. The transdermal delivery composition of claim77, wherein said delivered agent is selected from the group consistingof tramadol hydrochloride, fentanyl, metamizole, morphine sulphate,ketorolac tromethamine, hydrocodone, oxycodone, morprhine, loxoprofen,Capsaicin, and Boswellin.
 79. The transdermal delivery composition ofclaim 49, wherein said delivered agent is an NSAID.
 80. The transdermaldelivery composition of claim 79, wherein said non-steroidal antiinflammatory drug (NSAID) is selected from the group consisting ofibuprofen (2-(isobutylphenyl)-propionic acid); methotrexate (N-[4-(2, 4diamino 6-pteridinyl-methyl]methylamino]benzoyl)-L-glutamic acid);aspirin (acetylsalicylic acid); salicylic acid; diphenhydramine(2-(diphenylmethoxy)-NN-dimethylethylamine hydrochloride); naproxen(2-naphthaleneacetic acid, 6-methoxy-9-methyl-, sodium salt, (−));phenylbutazone (4-butyl-1,2-diphenyl-3,5-pyrazolidinedione);sulindac-(2)-5-fluoro-2-methyl-1-[[p-(methylsulfinyl)phenyl]methylene-]-1H-indene-3-aceticacid; diflunisal (2′,4′,-difluoro-4-hydroxy-3-biphenylcarboxylic acid;piroxicam(4-hydroxy-2-methyl-N-2-pyridinyl-2H-1,2-benzothiazine-2-carboxamide1,1-dioxide, an oxicam; indomethacin(1-(4-chlorobenzoyl)-5-methoxy-2-methyl-H-indole-3-acetic acid);meclofenamate sodium (N-(2,6-dichloro-m-tolyl) anthranilic acid, sodiumsalt, monohydrate); ketoprofen (2-(3-benzoylphenyl)-propionic acid;tolmetin sodium (sodium 1-methyl-5-(4-methylbenzoyl-1H-pyrrole-2-acetatedihydrate); diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneaticacid, monosodium salt); hydroxychloroquine sulphate(2-{[4-[(7-chloro-4-quinolyl)amino]pentyl]ethylamino}ethanol sulfate(1:1); penicillamine (3-mercapto-D-valine); flurbiprofen([1,1-b]phenyl]-4-acetic acid, 2-fluoro-alphamethyl-, (+−)); cetodolac(1-8-diethyl-13,4,9, tetrahydropyrano-[3-4-13]indole-1-acetic acid;mefenamic acid (N-(2,3-xylyl)anthranilic acid; and diphenhydraminehydrochloride (2-diphenyl methoxy-N,N-di-methylethamine hydrochloride).81. The transdermal delivery composition of claim 49, wherein thedelivered agent is an anti-inflammatory compound selected from the groupconsisting of hydrocortisone, prednisolone, triamcinolone, cortisone andpiroxicam.
 82. The transdermal delivery composition of claim 49, whereinthe delivered agent is an anti-infective compound selected from thegroup consisting of amoxicillin, clavulanate potassium, itraconazole,acyclovir, fluconazole, terbinafine hydrochloride, erythromycinethylsuccinate, acetyl sulfisoxazole, penicillin V, cephalexin,erythromycin, azithromycin, tetracycline, ciproflaxin, gentamycin,sulfathiazole, nitrofurantoin, norfloxacin, flumequine, and ibafloxacin,metronidazole, nystatin, lamivudine, indinavir sulfate, and stavudine.83. The transdermal delivery composition of claim 49, wherein thedelivered agent is metformin, metformin hydrochloride or acarbose. 84.The transdermal delivery composition of claim 49, wherein the deliveredagent is an immunogen.
 85. The transdermal delivery composition of claim49, wherein the immunogen comprises a polynucleotide.
 86. Thetransdermal delivery composition of claim 49 wherein the delivered agentis an immune response modifier.
 87. The transdermal delivery compositionof claim 86, wherein the immune response modifier is selected from thegroup consisting of purine derivatives, adenine derivatives,cyclosporine, and CpGs.
 88. The transdermal delivery composition ofclaim 49, wherein the delivered agent is selected from the groupconsisting of zileuton, captopril, and lisinopril.
 89. The transdermaldelivery composition of claim 49, wherein the delivered agent isselected from the group consisting of ergotamine, melatonin,sumatriptan, zolmitriptan, and rizatriptan.
 90. The transdermal deliverycomposition of claim 49, wherein the delivered agent is selected fromthe group consisting of zolpidem, zolpidem tartrate, triazolam, andhycosine butylbromide.
 91. The transdermal delivery composition of claim49, wherein the delivered agent is selected from the group consisting ofiohexyl, technetium, TC99M, sestamibi, iomeprol, gadodiamide, ioversol,and iopromide.
 92. The transdermal delivery composition of claim 49,wherein the delivered agent is selected from the group consisting ofalsactide, americium, betazole, histamine, mannitol, metyrapone,petagastrin, phentolamine, radioactive B₁₂, gadodiamide, gadopenteticacid, gadoteridol, perflubron, cyclosporine, sildenafil citrate,paclitaxel, ritonavir, and saquinavir.
 93. A transdermal delivery devicecomprising the transdermal delivery composition of claim 1, wherein saiddevice comprises: a removable cartridge, wherein said removablecartridge is configured to contain said transdermal deliverycomposition, and wherein said removable cartridge comprises a movablewall and a one-way valve; a body portion, said body portion beingadaptable to receive the removable cartridge, wherein the body portioncomprises an internal wall having an aperture configured to communicatewith the one-way valve; a movable member, wherein said movable memberpartially defines a dosing chamber along with the internal wall, andwherein movement of said movable member alters the volume of said dosingchamber; a plunger, said plunger being actuatable between a firstposition wherein said plunger interacts with the movable wall of saidremovable cartridge, and a second position, wherein said plunger isbiased to be in said second position; a non-invasive applicator; and aslidable member attached to said plunger, said slidable member beingactuatable between a first position and a second position correspondingto the first and second positions of said plunger, wherein when saidslidable member is in said first position, fluid communication betweenthe one-way valve and the dosing chamber is permitted, and wherein whensaid slidable member is in said second position, fluid communicationbetween said dosing chamber and the non-invasive applicator ispermitted.
 94. The transdermal delivery device of claim 93, furthercomprising a rotatable shaft, wherein said rotatable shaft has athreaded portion extending through a threaded aperture in the moveablemember, whereby rotation of the rotatable shaft results translation ofthe movable member in the direction of the axis of the shaft.
 95. Thetransdermal delivery device of claim 94, wherein the dosing chamber hasa non-circular cross-section, and wherein the movable member comprises alayer of deformable material along the edges in contact with the wallsof the dosing chamber.
 96. The transdermal delivery device of claim 94,wherein the rotatable shaft extends through a portion of the moveablemember away from the center of the upper surface of the movable member.97. The transdermal delivery device of claim 93, wherein thenon-invasive applicator comprises a roll-on applicator.
 98. Thetransdermal delivery device of claim 93, wherein the slidable member isoperably connected to the plunger via a spring.
 99. The transdermaldelivery device of claim 93, wherein the plunger is biased to be in saidsecond position via a spring connected to said body portion.
 100. Thetransdermal delivery device of claim 93, wherein the plunger comprises atoothed surface, said toothed surface configured to interact with alocking member to prevent return of said plunger to the second position.101. The transdermal delivery device of claim 100, wherein the lockingmember is operably connected to a release button, whereby pressing therelease button moves the locking member so as to permit return of theplunger to the second position.
 102. A transdermal delivery device fordispensing a measured quantity of the transdermal delivery compositionof claim 1, the device comprising: a reservoir, said reservoircomprising a movable wall and a one-way valve, wherein said movable wallcan be actuated to result in fluid flow through the one-way valve; aplunger, said plunger being actuatable between a first position, whereinsaid plunger is not in contact with the movable wall of said reservoir,and a second position, wherein said plunger interacts with the movablewall of the reservoir and further translation of the plunger in thedirection of the movable wall results in fluid flow through the one-wayvalve of the reservoir; and a dosing member, said dosing member beingmovable between a plurality of positions, wherein each of said pluralityof positions corresponds to a given quantity of transdermal deliverycomposition to be dispensed. 103-110. (canceled)
 111. A method ofproviding a delivered agent to a mammal in need thereof, comprising:providing the transdermal delivery device of claim 93; obtaining anamount of transdermal delivery system comprising said delivered agentfrom said transdermal delivery device; and contacting said mammal withsaid transdermal delivery system.
 112. A method of providing a deliveredagent to a mammal in need thereof, comprising: obtaining an amount oftransdermal delivery system of claim 1 comprising said delivered agentfrom said transdermal delivery device; contacting said mammal with saidtransdermal delivery system; and monitoring said mammal for the effectof said delivered agent.
 113. A method of providing a delivered agent toa mammal in need thereof, comprising: obtaining an amount of transdermaldelivery system of claim 49 comprising said delivered agent from saidtransdermal delivery device; contacting said mammal with saidtransdermal delivery system; and monitoring said mammal for the effectof said delivered agent.
 114. A transdermal delivery device comprisingthe transdermal delivery composition of claim 49, wherein said devicecomprises: a removable cartridge, wherein said removable cartridge isconfigured to contain said transdermal delivery composition, and whereinsaid removable cartridge comprises a movable wall and a one-way valve; abody portion, said body portion being adaptable to receive the removablecartridge, wherein the body portion comprises an internal wall having anaperture configured to communicate with the one-way valve; a movablemember, wherein said movable member partially defines a dosing chamberalong with the internal wall, and wherein movement of said movablemember alters the volume of said dosing chamber; a plunger, said plungerbeing actuatable between a first position wherein said plunger interactswith the movable wall of said removable cartridge, and a secondposition, wherein said plunger is biased to be in said second position;a non-invasive applicator; and a slidable member attached to saidplunger, said slidable member being actuatable between a first positionand a second position corresponding to the first and second positions ofsaid plunger, wherein when said slidable member is in said firstposition, fluid communication between the one-way valve and the dosingchamber is permitted, and wherein when said slidable member is in saidsecond position, fluid communication between said dosing chamber and thenon-invasive applicator is permitted.
 115. The transdermal deliverydevice of claim 114, further comprising a rotatable shaft, wherein saidrotatable shaft has a threaded portion extending through a threadedaperture in the moveable member, whereby rotation of the rotatable shaftresults translation of the movable member in the direction of the axisof the shaft.
 116. The transdermal delivery device of claim 114, whereinthe dosing chamber has a non-circular cross-section, and wherein themovable member comprises a layer of deformable material along the edgesin contact with the walls of the dosing chamber.
 117. The transdermaldelivery device of claim 115, wherein the rotatable shaft extendsthrough a portion of the moveable member away from the center of theupper surface of the movable member.
 118. The transdermal deliverydevice of claim 114, wherein the non-invasive applicator comprises aroll-on applicator.
 119. The transdermal delivery device of claim 114,wherein the slidable member is operably connected to the plunger via aspring.
 120. The transdermal delivery device of claim 114, wherein theplunger is biased to be in said second position via a spring connectedto said body portion.
 121. The transdermal delivery device of claim 114,wherein the plunger comprises a toothed surface, said toothed surfaceconfigured to interact with a locking member to prevent return of saidplunger to the second position.
 122. The transdermal delivery device ofclaim 121, wherein the locking member is operably connected to a releasebutton, whereby pressing the release button moves the locking member soas to permit return of the plunger to the second position.
 123. Atransdermal delivery device for dispensing a measured quantity of thetransdermal delivery composition of claim 49, the device comprising: areservoir, said reservoir comprising a movable wall and a one-way valve,wherein said movable wall can be actuated to result in fluid flowthrough the one-way valve; a plunger, said plunger being actuatablebetween a first position, wherein said plunger is not in contact withthe movable wall of said reservoir, and a second position, wherein saidplunger interacts with the movable wall of the reservoir and furthertranslation of the plunger in the direction of the movable wall resultsin fluid flow through the one-way valve of the reservoir; and a dosingmember, said dosing member being movable between a plurality ofpositions, wherein each of said plurality of positions corresponds to agiven quantity of transdermal delivery composition to be dispensed.